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Conventional agricultural practices based on the application of synthetic fertilizers are increasingly considered as unsustainable. Under a forecasted scenario of drought for the next decades, there is a global demand for innovative and sustainable approaches to ameliorate plant performance. Here, encapsulating beneficial microbes (BMs) to promote plant growth is gaining attention. This study evaluates bacterial encapsulation using polymeric beads of alginate, testing the survival of Pseudomonas libanensis TR1 stored up to 90 days. Produced beads were subjected to different treatments (fresh, air-dried and pulverized), which resulted in a variable size range (1200–860 µm). After storage, bacterial viability was maintained, and air-dried beads displayed a higher number of colony-forming units (2 × 107). Then, a glasshouse experiment investigated the drought resistance (plant growth, biomass, and photosynthetic responses) of Vigna unguiculata plants inoculated with these alginate beads. After 10 days of complete water restriction, turgidity and relative water content of V. unguiculata were still high under drought stress (> 80%). Leaf and root growth and biomass did not evidence significant changes after water restriction even after P. libanensis inoculation. Plant photosynthetic parameters (stomatal conductance, net photosynthetic rate, leaf CO2 concentration, or Fv'/Fm') were slightly affected due to inoculation but the level of stress-induced minimal plant responses. In our experiment, water restriction might have been insufficient to downregulate photosynthetic efficiency and reduce plant growth, limiting our understanding of the role of P. libanensis inoculation in alleviating drought stress in V. unguiculata, but highlighting the important relationship between the stress level and agricultural benefits of using encapsulated BMs.

Ectomycorrhizal fungi (ECMF) play an important role in forest ecosystems, often mitigating stress factors and increasing seedling performance. The aim of this study was to investigate the effects of a nursery inoculation on Pinus pinaster growth and on the fungal communities established when reforesting burned areas. Inoculated P. pinaster saplings showed 1.5-fold higher stem height than the non-inoculated controls after a 5 year growth period, suggesting that fungal inoculation could potentiate tree growth in the field. Ordination analysis revealed the presence of different ECMF communities on both plots. Among the nursery-inoculated fungi, Laccaria sp., Rhizopogon sp., Suillus bovinus and Pisolithus sp. were detected on inoculated Pinus saplings on both sampling periods, indicating that they persisted after field establishment. Other fungi were also detected in the inoculated plants. Phialocephala sp. was found on the first assessment, while Terfezia sp. was detected on both sampling periods. Laccaria sp. and Rhizopogon sp. were identified in the control saplings, belonging however to different species than those found in the inoculated plot. Inocybe sp., Thelephora sp. and Paxillus involutus were present on both sampling periods in the non-inoculated plots. The results suggest that ECMF inoculation at nursery stage can benefit plant growth after transplantation to a post-fire site and that the inoculated fungi can persist in the field. This approach has great potential as a biotechnological tool to aid in the reforestation of burned areas.

Diflubenzuron (DFB) is an insecticide commonly used to control forest pests. The objectives of this study were to assess the effect of diflubenzuron on the development of Pinus pinaster seedlings and Pisolithus tinctorius under laboratory conditions and to study the possible protective role of this ectomycorrhizal fungus against the effects of diflubenzuron. In vitro experiments revealed that diflubenzuron inhibited fungal growth at all tested concentrations (0.01, 0.1, 1, 10 and 100 mg L(-1)). Root growth was inhibited at the two highest diflubenzuron concentrations. The activity of the antioxidant defence system of non-inoculated P. pinaster increased at 1 and 10 mg DFB kg(-1) substrate, and inoculation increased the threshold to the highest concentration. The protective role of the ectomycorrhizal fungus was seen in the increase of CAT activity. This study revealed that despite causing no mortality, diflubenzuron has the ability to cause sub-lethal damage to P. pinaster. The disproportionate use of this insecticide may lead to higher amounts of its residues in soil and the biosphere, endangering trees, fungi and their symbiosis.

A field survey of the arbuscular mycorrhizal status of herbaceous plant species was conducted in a highly alkaline anthropogenic sediment resulting from the disposal of waste from an acetylene and polyvinyl chloride factory. Most plant species found at the site were mycorrhizal and the dominant mycotrophic plant species was Conyza bilbaoana. Fungal species richness was assessed by identification of spores extracted from the sediment and from continuously propagated trap pot cultures. All of the six species of arbuscular mycorrhizal fungi (AMF) found were from the genus Glomus. Glomus intraradices and G. mosseae were found in field-collected sediment samples and also occurred most frequently in trap cultures. To test the symbiotic effectiveness of these two fungi, seedlings of C. bilbaoana were inoculated with either native G. intraradices BEG163 or G. mosseae BEG198 and non-native G. intraradices BEG75 or G. mosseae BEG25 isolates in sterile and non-sterile sediment collected from the study site. All four isolates were able to colonise C. bilbaoana. However, AMF native to the target sediments were generally more effective than the non-native fungi in promoting plant establishment and growth under highly alkaline conditions. The non-native G. intraradices was, however, more effective than the non-native G. mosseae. The results of this study suggest the use of adapted AMF as inoculants for phytorestoration of alkaline anthropogenic-stressed sediments.