Priming of Plant Growth Promotion by Volatiles of Root-Associated Microbacterium spp.

Volatile compounds produced by plant-associated microorganisms represent a diverse resource to promote plant growth and health. Here, we investigated the effect of volatiles from root-associated Microbacterium species on plant growth and development. Volatiles of eight strains induced significant increases in shoot and root biomass of Arabidopsis but differed in their effects on root architecture. Microbacterium strain EC8 also enhanced root and shoot biomass of lettuce and tomato. Biomass increases were also observed for plants exposed only briefly to volatiles from EC8 prior to transplantation of the seedlings to soil. These results indicate that volatiles from EC8 can prime plants for growth promotion without direct and prolonged contact. We further showed that the induction of plant growth promotion is tissue specific; that is, exposure of roots to volatiles from EC8 led to an increase in plant biomass, whereas shoot exposure resulted in no or less growth promotion. Gas chromatography-quadrupole time of flight mass spectometry (GC-QTOF-MS) analysis revealed that EC8 produces a wide array of sulfur-containing compounds, as well as ketones. Bioassays with synthetic sulfur volatile compounds revealed that the plant growth response to dimethyl trisulfide was concentration-dependent, with a significant increase in shoot weight at 1 µM and negative effects on plant biomass at concentrations higher than 1 mM. Genome-wide transcriptome analysis of volatile-exposed Arabidopsis seedlings showed upregulation of genes involved in assimilation and transport of sulfate and nitrate. Collectively, these results show that root-associated Microbacterium primes plants, via the roots, for growth promotion, most likely via modulation of sulfur and nitrogen metabolism.IMPORTANCE In the past decade, various studies have described the effects of microbial volatiles on other (micro)organisms in vitro, but their broad-spectrum activity in vivo and the mechanisms underlying volatile-mediated plant growth promotion have not been addressed in detail. Here, we revealed that volatiles from root-associated bacteria of the genus Microbacterium can enhance the growth of different plant species and can prime plants for growth promotion without direct and prolonged contact between the bacterium and the plant. Collectively, these results provide new opportunities for sustainable agriculture and horticulture by exposing roots of plants only briefly to a specific blend of microbial volatile compounds prior to transplantation of the seedlings to the greenhouse or field. This strategy has no need for large-scale introduction or root colonization and survival of the microbial inoculant.

Do fungicides used to control Rhizoctonia solani impact the non-target arbuscular mycorrhizal fungus Rhizophagus irregularis?

There is growing evidence that the application of biocontrol organisms (e.g., Pseudomonas and Bacillus spp., arbuscular mycorrhizal fungi-AMF) is a feasible option to reduce incidence of plant pathogens in an integrated control strategy. However, the utilization of these microorganisms, in particular AMF, may be threatened by the application of fungicides, a widely-used measure to control Rhizoctonia solani in various crops among which potato. Prior to their application, it is thus important to determine the impact of fungicides on AMF. The present study investigated, under in vitro controlled conditions, the impact of azoxystrobin (a systemic broad-spectrum fungicide), flutolanil (a systemic Basidiomycota-specific fungicide), and pencycuron (a contact Rhizoctonia-specific fungicide) and their respective formulations (Amistar, Monarch, and Monceren) on the growth and development of the AMF Rhizophagus irregularis MUCL 41833 (spore germination, root colonization, extraradical mycelium development, and spore production) at doses used to control R. solani. Results demonstrated that azoxystrobin and its formulation Amistar, at threshold values for R. solani control (estimated by the half maximal inhibitory concentration, IC50, on a dry weight basis), did not affect spore germination and potato root colonization by R. irregularis, while the development of extra-radical mycelium and spore production was reduced at 10 times the threshold value. Flutolanil and its formulation Monarch at threshold value did not affect spore germination or extra-radical development but decreased root colonization and arbuscule formation. At threshold value, pencycuron and its formulation Monceren, did not affect spore germination and intra- or extraradical development of R. irregularis. These results suggest that azoxystrobin and pencycuron do not affect the AMF at threshold concentrations to control R. solani in vitro, while flutolanil (as formulation) impacts the intraradical phase of the fungus. These fungicides and R. irregularis thus have the potential to be used in parallel against Rhizoctonia disease in potato.

Radiocesium transfer between Medicago truncatula plants via a common mycorrhizal network.

Common mycorrhizal networks of arbuscular mycorrhizal fungi have been reported to transfer cesium between plants. However, a direct hyphae-mediated transfer (via cytoplasm/protoplasm) cannot be distinguished from an indirect transfer. Indeed, cesium released by the roots of the donor plant can be taken up by the receiver plant or fungal hyphae. In the present study, Medicago truncatula plants were connected by a common mycorrhizal network and Prussian Blue (ammonium-ferric-hexacyano ferrate) was added in the growth medium to adsorb the released radiocesium. A direct transfer of radiocesium to roots and shoots of the receiver plant was clearly demonstrated for the first time. Even though this transfer was quantitatively low, it suggested that shared mycorrhizal networks could contribute to the redistribution of this radionuclide in the environment, which otherwise would be restricted both in time and space. This finding may also help to understand the behaviour of its chemical analogue, potassium.

Development of an autotrophic culture system for the in vitro mycorrhization of potato plantlets.

An autotrophic culture system was developed for the in vitro mycorrhization of potato plantlets. Roots of micropropagated plantlets were associated to an arbuscular mycorrhizal fungus under in vitro conditions, while shoots developed under open air conditions. Several thousand spores, an extensive extraradical mycelium and an abundant root colonization were obtained. Spores were able to colonize new plantlets under the same conditions. These results support the capacity of the autotrophic culture system to continuously culture arbuscular mycorrhizal fungi and may serve as a powerful tool to investigate various aspects of the symbiosis for which a source-sink relationship and photosynthetic active tissues are necessary.

Effects of arbuscular mycorrhizal fungi on the root uptake and translocation of radiocaesium.

Because mycorrhizal fungi are intimately associated with plant roots, their importance in radionuclide (RN) recycling and subsequent dispersion into the biosphere has received an increasing interest. Recently, the capacity of arbuscular mycorrhizal fungi to take up and translocate radiocaesium to their host was demonstrated. However, the relative contribution of these processes in comparison to the ones of roots remains unknown. Here, the respective contributions of the hyphae of a Glomus species and the transformed carrot (Daucus carota L.) roots on radiocaesium uptake and translocation were compared and quantified. We observed that radiocaesium uptake by hyphae was significantly lower as compared to that of the roots, while the opposite was noted for radiocaesium translocation/uptake ratio. We also observed that the intraradical fungal structures might induce a local accumulation of radiocaesium and concurrently reduce its translocation within mycorrhizal roots. We believe that intraradical fungal structures might induce the down-regulation of radiocaesium channels involved in the transport processes of radiocaesium towards the xylem.

Polyaromatic hydrocarbons impair phosphorus transport by the arbuscular mycorrhizal fungus Rhizophagus irregularis.

Phosphate uptake by plant roots is mainly mediated by arbuscular mycorrhizal fungi (AMF). However, the impact on phosphorus (P) transport of polycyclic aromatic hydrocarbons (PAH), persistent organic pollutants widely found in altered soils, is not known up today. Here, we monitored the Rhizophagus irregularis fungal growth and the fungal P transport ability from the extraradical mycelium to the host transformed chicory roots in the presence of anthracene and benzo[a]pyrene (B[a]P) and the combination of both PAH, under in vitro conditions. Firstly, our findings showed that PAH have detrimental effect on the fungal growth. The combination of both PAH was more toxic than each of the PAH individually due to synergistic effects. Secondly, PAH affected the P transport by the fungus from the medium to the roots. This was evidenced by either the decrease in (33)P quantity transported in the roots as well as the decrease in acid phosphatase activity in the mycorrhizal roots. Moreover, the fungal alkaline phosphatase activities remained constant in the extraradical mycelium as well as in the roots in the absence and in the presence of PAH. The GintPT and GiALP (encoding a P transporter and an alkaline phosphatase respectively) gene expressions were also found to be similar in the extraradical mycelium treated with PAH or not (control). These findings suggested that the P uptake by R. irregularis was not affected by PAH but probably the transport from the extraradical mycelium to the intraradical mycelium.

Arbuscular mycorrhizal fungi decrease radiocesium accumulation in Medicago truncatula.

The role of arbuscular mycorrhizal fungi (AMF) in plant radiocesium uptake and accumulation remains ambiguous. This is probably due to the presence of other soil microorganisms, the variability of soil characteristics and plant nutritional status or the availability of its chemical analogue, potassium (K). Here, we used an in vitro culture system to study the impact of increased concentration of K on radiocesium accumulation in non K-starved mycorrhizal and non-mycorrhizal Medicago truncatula plants. In the presence of AMF radiocesium uptake decreased regardless of the concentration of K, and its translocation from root to shoot was also significantly lower. Potassium also reduced the accumulation of radiocesium in plants but to a lesser extent than mycorrhization, and without any effect on translocation. These results suggest that AMF in combination with K can play a key role in reducing radiocesium uptake and its subsequent translocation to plant shoots, thereby representing good potential for improved phytomanagement of contaminated areas.

Effect of potassium and phosphorus on the transport of radiocesium by arbuscular mycorrhizal fungi.

Potassium, a chemical analogue of cesium, and phosphorus, an essential macronutrient transported by arbuscular mycorrhizal fungi (AMF), have been suggested to influence the transport of radiocesium by AMF. However, no study investigated the effects of increasing concentrations of both elements on the importance of this transport. Here, the arbuscular mycorrhizal-plant (AM-P) in vitro culture system associating Medicago truncatula plantlets with Glomus intraradices was used to evaluate this effect. Using three concentrations of K (0, 1, 10 mM) and two concentrations of P (30 and 3000 microM) added to a compartment only accessible to the AMF, we demonstrated that K and P individually and in combination significantly influenced radiocesium transport by AMF. Whilst increased concentration of K decreased the amount of radiocesium transported, the opposite was observed for P. Although the exact mechanisms involved need to be assessed, both elements were identified as important factors influencing the transport of radiocesium by AMF.

Extraradical mycelium of the arbuscular mycorrhizal fungus Glomus lamellosum can take up, accumulate and translocate radiocaesium under root-organ culture conditions.

Radiocaesium enters the food chain when plants absorb it from soil, in a process that is strongly dependent on soil properties and plant and microbial species. Among the microbial species, arbuscular mycorrhizal (AM) fungi are obligate symbionts that colonize the root cortex of many plants and develop an extraradical mycelial (ERM) network that ramifies in the soil. Despite the well-known involvement of this ERM network in mineral nutrition and uptake of some heavy metals, only limited data are available on its role in radiocaesium transport in plants. We used root-organ culture to demonstrate that the ERM of the AM fungus Glomus lamellosum can take up, possibly accumulate and unambiguously translocate radiocaesium from a 137Cs-labelled synthetic root-free compartment to a root compartment and within the roots. The accumulation of 137Cs by hyphae in the root-free compartment may be explained by sequestration in the hyphae or by a bottleneck effect resulting from a limited number of hyphae crossing the partition between the two compartments. Uptake and translocation resulted from the incorporation of 137Cs into the fungal hyphae, as no 137Cs was detected in mycorrhizal roots treated with formaldehyde. The importance of the translocation process was indicated by the correlation between 137Cs measured in the roots and the total hyphal length connecting the roots with the labelled compartment. 137Cs may be translocated via a tubular vacuolar system or by cytoplasmic streaming per se.