Late blight caused by Phytophthora infestans is considered as the most devastating disease of potato and is a re-emerging problem worldwide. Current late blight control practices rely mostly on synthetic fungicides or copper-based products, but growing awareness of the negative impact of these compounds on the environment has led to the search for alternative control measures. A collection of Pseudomonas strains isolated from both the rhizosphere and the phyllosphere of potato was recently characterized for in vitro protective effects against P. infestans. In the present study, we used a leaf disk assay with three different potato cultivars to compare the disease inhibition capacity of nine selected Pseudomonas strains when applied alone or in all possible dual and triple combinations. Results showed a strong cultivar effect and identified strains previously thought to be inactive based on in vitro assays as the best biocontrol candidates. One strain was much more active alone than in combination with other strains, while two other strains provided significantly better protection in dual combination than when applied alone. A subset of five strains was then further selected to determine their mutual influence on each other’s survival and growth, as well as to characterize their activity against P. infestans in more details. This revealed that the two strains whose dual combination was particularly efficient were only weakly interfering with each other’s growth and had complementary modes of action. Our results highlight the potential to harness the crop’s native rhizosphere and phyllosphere microbiome through re-assembling strains with differing modes of action into small communities, thereby providing more consistent protection than with the application of single strains. We consider this as a first step toward more elaborate microbiome management efforts, which shall be integrated into global strategies for sustainable control of potato late blight.

The plant kingdom represents a prominent biodiversity island for microbes that associate with the below- or aboveground organs of vegetal species. Both the root and the leaf represent interfaces where dynamic biological interactions influence plant life. Beside well-studied communication strategies based on soluble compounds and protein effectors, bacteria were recently shown to interact both with host plants and other microbial species through the emissions of volatile organic compounds (VOCs). Focusing on the potato late blight-causing agent Phytophthora infestans, this work addresses the potential role of the bacterial volatilome in suppressing plant diseases. In a previous study, we isolated and identified a large collection of strains with anti-Phytophthora potential from both the phyllosphere and the rhizosphere of potato. Here we report the characterization and quantification of their emissions of biogenic volatiles, comparing 16 Pseudomonas strains differing in i) origin of isolation (phyllosphere vs. rhizosphere), ii) in vitro inhibition of P. infestans growth and sporulation behavior, and iii) protective effects against late blight on potato leaf discs. We systematically tested the pharmacological inhibitory activity of core and strain-specific single compounds against P. infestans mycelial growth and sporangial behavior in order to identify key effective candidate molecules present in the complex natural VOCs blends. We envisage the plant bacterial microbiome as a reservoir for functional VOCs and establish the basis for finding the primary enzymatic toolset that enables the production of active components of the volatile bouquet in plant-associated bacteria. Comprehension of these functional interspecies interactions will open perspectives for the sustainable control of plant diseases in forthcoming agriculture.

Late blight, caused by the oomycete Phytophthora infestans, is the most devastating disease of potato. In organic farming, late Late blight, caused by the oomycete Phytophthora infestans, is the most devastating disease of potato. In organic farming, late blight is controlled by repeated applications of copper-based products, which negatively impact the environment. To find alternative solutions for late blight management, we have previously isolated a large collection of bacteria from the phyllosphere and the rhizosphere of potatoes. Here we report the antagonistic potential of these strains when co-cultivated with P. infestans as well as with other potato pathogens. We then focused on three Pseudomonas strains and compared their protective impact against late blight to that of well-known biocontrol strains in planta using a high-throughput leaf disc assay with automated picture analysis. When sprayed on the leaves of potatoes in the greenhouse, the strains were able to survive for at least 15 days. Under field conditions, populations decreased faster but all tested strains could still be retrieved after 8 days. The most active strain in vitro, P. chlororaphis R47, was also the best protectant on leaf discs from plants grown in the greenhouse experiment, but its protection potential could not be verified in the field due to unfavourable infection conditions. However, its protective effect against P. infestans in planta, its survival in the phyllosphere as well as its ability to colonise the potato rhizosphere in very high population densities, suggest a potential for field application, e.g. in the form of tuber treatment or leaf spray.

Late blight caused by the oomycete Phytophthora infestans constitutes the greatest threat to potato production worldwide. Considering the increasing concerns regarding the emergence of novel fungicide-resistant genotypes and the general demand for reducing inputs of synthetic and copper-based fungicides, the need for alternative control methods is acute. Several bacterial antagonists have shown anti-Phytophthora effects during in vitro and greenhouse experiments. We report the effects of three Pseudomonas strains recovered from field-grown potatoes against a collection of P. infestans isolates assembled for this study. The collection comprised 19 P. infestans isolates of mating types A1 and A2 greatly varying in fungicide resistance and virulence profiles as deduced from leaf disc experiments on Black’s differential set. The mycelial growth of all P. infestans isolates was fully inhibited when co-cultivated with the most active Pseudomonas strain (R47). Moreover, the isolates reacted differently to exposure to the less active Pseudomonas strains (S19 and R76). Leaf disc infection experiments with six selected P. infestans isolates showed that four of them, including highly virulent and fungicide-resistant ones, could be efficiently controlled by different potato-associated Pseudomonas strains. [ ] Copyright © 2019 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Aim: Recent studies have shown that plants harbor complex bacterial communities, the so-called “microbiome”. We are only beginning to unravel the origin of these bacterial plant inhabitants, their community structure and their roles, which, in analogy to the gut microbiome, are likely to be of essential nature. The aim of this work was to analyze the abundance and diversity of the cultivable members of the bacterial microbiome living in and on the leaves of grapevine, and to identify microbiome members with putative plant-protective activities against phytopathogenic organisms. Methods and results: Grapevine leaves were sampled three times during the growing season at one location and used to quantify the abundance and isolate representative members of the epiphytic and endophytic bacterial communities. Results were compared for three grapevine cultivars: Pinot noir, Chasselas and Solaris. Greater bacterial abundance and diversity was observed among epiphytes than among endophytes. Leaf imprints revealed a higher colonization density for the upper than for the lower surface of leaves. A high relative frequency of strains belonging to the genera Staphylococcus and Bacillus was observed, especially in the endophytic communities. Conclusions: The first results of this ongoing study led to the conclusion that epiphytic bacteria of the cultivable grapevine microbiome were more abundant and diverse than endophytic bacteria. A tendency towards more abundant bacteria in the resistant variety Solaris than in the susceptible varieties Chasselas and Pinot noir contrasted with a higher diversity of epiphytic bacteria in the Pinot noir variety. A trend for higher frequency of strains showing antagonistic activity towards the grapevine pathogen Botrytis cinerea among isolates from the resistant variety Solaris could indicate a putative contribution of microbiome bacteria to this resistant phenotype, even though this remains to be confirmed. Significance and impact of the study: This study constitutes a first step in characterizing the endo- and epiphytic cultivable bacterial communities of three grapevine varieties grown at the same location. Further, presently ongoing studies shall reveal i) the true complexity of these leaf-associated communities through cultivation-independent, next generation sequencing techniques, and ii) their potential as a source of biofungicidal agents. In this last point, the contribution of the emission of volatile organic compounds (VOCs) as a newly discovered type of bacterial weapon against fungal and oomycetal pathogens shall be quantified.

Bacterial volatiles play important roles in mediating beneficial interactions between plants and their associated microbiota. Despite their relevance, bacterial volatiles are mostly studied under laboratory conditions, although these strongly differ from the natural environment bacteria encounter when colonizing plant roots or shoots. In this work, we ask the question whether plant-associated bacteria also emit bioactive volatiles when growing on plant leaves rather than on artificial media. Using four potato-associated Pseudomonas, we demonstrate that potato leaves offer sufficient nutrients for the four strains to grow and emit volatiles, among which 1-undecene and Sulfur compounds have previously demonstrated the ability to inhibit the development of the oomycete Phytophthora infestans, the causative agent of potato late blight. Our results bring the proof of concept that bacterial volatiles with known plant health-promoting properties can be emitted on the surface of leaves and warrant further studies to test the bacterial emission of bioactive volatiles in greenhouse and field-grown plants.