The study of arbuscular mycorrhizal (AM) fungi and the AM symbiosis formed with host plant roots is complicated by the biotrophic and hypogeous nature of the mycobionts involved. To overcome this, several attempts have been made during the last three decades to obtain this symbiosis in vitro. The use of root-organ cultures has proved particularly successful. In this review, we describe the method by which root-organ cultures (transformed and nontransformed) have been obtained, together with the choice of host species, inoculation techniques, and culture media. We also outline the potential use of continuous cultures and cryopreservation of in vitro produced spores for long-term germ plasm storage. Furthermore, this review highlights the considerable impact that in vitro root-organ cultures have had on studies of AM fungal morphology, taxonomy, and phylogeny and how they have improved our understanding of the processes leading to root colonization and development of the extraradical mycelium. This is supported by a summary of some of the most important findings, regarding this symbiosis, that have been made at the physiological, biochemical, and molecular levels. We also summarize results from studies between AM fungi and certain pathogenic and nonpathogenic soil microorganisms. We describe some of the limitations of this in vitro system and propose diverse avenues of AM research that can now be undertaken, including the potential use of a similar system for ectomycorrhizal research.Key words: arbuscular mycorrhiza, root-organ cultures, Glomales, in vitro, root symbioses, source of inoculum, cryopreservation, intraradical and extraradical mycelium, mycorrhizosphere.

Cooperation is a prevalent phenomenon in nature and how it originates and maintains is a fundamental question in ecology. Many efforts have been made to understand cooperation between individuals in the same species, while the mechanisms enabling cooperation between different species are less understood. Here, we investigated under strict in vitro culture conditions if the exchange of carbon and phosphorus is pivotal to the cooperation between the arbuscular mycorrhizal fungus (AMF) Rhizophagus irregularis and the phosphate solubilizing bacterium (PSB) Rahnella aquatilis. We observed that fructose exuded by the AMF stimulated the expression of phosphatase genes in the bacterium as well as the rate of phosphatase release into the growth medium by regulating its protein secretory system. The phosphatase activity was subsequently increased, promoting the mineralization of organic phosphorus (i.e., phytate) into inorganic phosphorus, stimulating simultaneously the processes involved in phosphorus uptake by the AMF. Our results demonstrated for the first time that fructose not only is a carbon source, but also plays a role as a signal molecule triggering bacteria-mediated organic phosphorus mineralization processes. These results highlighted the molecular mechanisms by which the hyphal exudates play a role in maintaining the cooperation between AMF and bacteria.

Macrophomina phaseolina is a generalist soil-borne fungus present all over the world. It cause diseases such as stem and root rot, charcoal rot and seedling blight. Under high temperatures and low soil moisture, this fungus can cause substantial yield losses in crops such as soybean, sorghum and groundnut. The wide host range and high persistence of M. phaseolina in soil as microsclerotia make disease control challenging. Therefore, understanding the basis of the pathogenicity mechanisms as well as its interactions with host plants is crucial for controlling the pathogen. In this work, we aim to describe the general characteristics and pathogenicity mechanisms of M. phaseolina, as well as the hosts defense response. We also review the current methods and most promising forecoming ones to reach a responsible control of the pathogen, with minimal impacts to the environment and natural resources.

Many different cultivation techniques and inoculum products of the plant-beneficial arbuscular mycorrhizal (AM) fungi have been developed in the last decades. Soil- and substrate-based production techniques as well as substrate-free culture techniques (hydroponics and aeroponics) and in vitro cultivation methods have all been attempted for the large-scale production of AM fungi. In this review, we describe the principal in vivo and in vitro production methods that have been developed so far. We present the parameters that are critical for optimal production, discuss the advantages and disadvantages of the methods, and highlight their most probable sectors of application.

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.