Microbes to the Rescue – Exploring the Chemistry of Microbial Communication and Using it to Protect Plant Health

Plants are densely colonized by diverse microbial communities. These microbes, which provide important benefit to their host supporting its growth and health, interact with each other and with their host plant by exchanging chemical signals, among which volatile organic compounds (VOCs). This review presents some of our current research lines in the field of microbial VOCs, including their bioactivities on both plants and plant pathogens, and the abiotic and biotic factors influencing their emission. Understanding how VOCs emission is regulated in plant-associated microbes is one of the major challenges for both fundamental and translational aspects of this research field

Mining the volatilomes of plant-associated microbiota for new biocontrol solutions

Microbial lifeforms associated with land plants represent a rich source for crop growth- and health-promoting microorganisms and biocontrol agents. Volatile organic compounds (VOCs) produced by the plant microbiota have been demonstrated to elicit plant defenses and inhibit the growth and development of numerous plant pathogens. Therefore these molecules are prospective alternatives to synthetic pesticides and the determination of their bioactivities against plant threats could contribute to the development of control strategies for sustainable agriculture. In our previous study we investigated the inhibitory impact of volatiles emitted by Pseudomonas species isolated from a potato field against the late blight-causing agent Phytophthora infestans. Besides the well-documented emission of hydrogen cyanide, other Pseudomonas VOCs impeded P. infestans mycelial growth and sporangia germination. Current advances in the field support the emerging concept that the microbial volatilome contains unexploited, eco-friendly chemical resources that could help select for efficient biocontrol strategies and lead to a greener chemical disease management in the field

Microbial life in the grapevine: what can we expect from the leaf microbiome?

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

Airborne medicine: bacterial volatiles and their influence on plant health

Like most other eukaryotes, plants do not live alone but in close association with a diverse microflora. These plant‐associated microbes contribute to plant health in many different ways, ranging from modulation of hormonal pathways to direct antibiosis of plant pathogens. Over the last 15 yr, the importance of volatile organic compounds as mediators of mutualistic interactions between plant‐associated bacteria and their hosts has become evident. This review summarizes current knowledge concerning bacterial volatile‐mediated plant protection against abiotic and biotic stresses. It then discusses the translational potential of such metabolites or of their emitters for sustainable crop protection, the possible ways to harness this potential, and the major challenges still preventing us from doing so. Finally, the review concludes with highlighting the most pressing scientific gaps that need to be filled in order to enable a better understanding of: the molecular mechanisms underlying the biosynthesis of bacterial volatiles; the complex regulation of bacterial volatile emission in natural communities; the perception of bacterial volatiles by plants; and the modes of actions of bacterial volatiles on their host

Secretion activity of white lupin's cluster roots influences bacterial abundance, function and community structure

White lupin (Lupinus albus L. cv. Amiga) reacts to phosphate deficiency by producing cluster roots which exude large amounts of organic acids. The detailed knowledge of the excretion physiology of the different root parts makes it a good model plant to study plant-bacteria interaction. Since the effect of the organic acid exudation by cluster roots on the rhizosphere microflora is still poorly understood, we investigated the abundance, diversity and functions of bacteria associated with the cluster roots of white lupin, with special emphasis on the influence of root proximity (comparing root, rhizosphere soil and bulk soil fractions) and cluster root growth stages, which are characterized by different excretion activities. Plants were grown for five weeks in microcosms, in the presence of low phosphate concentrations, on acidic sand inoculated with a soil suspension from a lupin field. Plate counts showed that bacterial abundance decreased at the stage where the cluster root excretes high amounts of citrate and protons. In vitro tests on isolates showed that the frequencies of auxin producers were highest in juvenile and mature cluster roots and significantly decreased in senescent cluster roots. However, no significant difference in the frequency of auxin producers was found between cluster and non cluster roots. The diversity and structure of bacterial communities were investigated by DGGE of 16S rDNA and 16S rRNA. The diversity and community structure were mostly influenced by root proximity and, to a lesser extent, by cluster root stage. The richness of bacterial communities decreased with root proximity, whereas the proportion of active populations increased. The high citrate and proton excretion occurring at the mature stage of cluster roots had a strong impact on the structure and richness of the bacterial communities, both in the root and in the rhizosphere soi

Secretion activity of white lupin's cluster roots influences bacterial abundance, function and community structure

White lupin (Lupinus albus L. cv. Amiga) reacts to phosphate deficiency by producing cluster roots which exude large amounts of organic acids. The detailed knowledge of the excretion physiology of the different root parts makes it a good model plant to study plant-bacteria interaction. Since the effect of the organic acid exudation by cluster roots on the rhizosphere microflora is still poorly understood, we investigated the abundance, diversity and functions of bacteria associated with the cluster roots of white lupin, with special emphasis on the influence of root proximity (comparing root, rhizosphere soil and bulk soil fractions) and cluster root growth stages, which are characterized by different excretion activities. Plants were grown for five weeks in microcosms, in the presence of low phosphate concentrations, on acidic sand inoculated with a soil suspension from a lupin field. Plate counts showed that bacterial abundance decreased at the stage where the cluster root excretes high amounts of citrate and protons. In vitro tests on isolates showed that the frequencies of auxin producers were highest in juvenile and mature cluster roots and significantly decreased in senescent cluster roots. However, no significant difference in the frequency of auxin producers was found between cluster and non cluster roots. The diversity and structure of bacterial communities were investigated by DGGE of 16S rDNA and 16S rRNA. The diversity and community structure were mostly influenced by root proximity and, to a lesser extent, by cluster root stage. The richness of bacterial communities decreased with root proximity, whereas the proportion of active populations increased. The high citrate and proton excretion occurring at the mature stage of cluster roots had a strong impact on the structure and richness of the bacterial communities, both in the root and in the rhizosphere soi

Production of Bioactive Volatiles by Different Burkholderia ambifaria Strains

Increasing evidence indicates that volatile compounds emitted by bacteria can influence the growth of other organisms. In this study, the volatiles produced by three different strains of Burkholderia ambifaria were analysed and their effects on the growth of plants and fungi, as well as on the antibiotic resistance of target bacteria, were assessed. Burkholderia ambifaria emitted highly bioactive volatiles independently of the strain origin (clinical environment, rhizosphere of pea, roots of maize). These volatile blends induced significant biomass increase in the model plant Arabidopsis thaliana as well as growth inhibition of two phytopathogenic fungi (Rhizoctonia solani and Alternaria alternata). In Escherichia coli exposed to the volatiles of B. ambifaria, resistance to the aminoglycoside antibiotics gentamicin and kanamycin was found to be increased. The volatile blends of the three strains were similar, and dimethyl disulfide was the most abundant compound. Sulfur compounds, ketones, and aromatic compounds were major groups in all three volatile profiles. When applied as pure substance, dimethyl disulfide led to increased plant biomass, as did acetophenone and 3-hexanone. Significant fungal growth reduction was observed with high concentrations of dimethyl di- and trisulfide, 4-octanone, S-methyl methanethiosulphonate, 1-phenylpropan-1-one, and 2-undecanone, while dimethyl trisulfide, 1-methylthio-3-pentanone, and o-aminoacetophenone increased resistance of E. coli to aminoglycosides. Comparison of the volatile profile produced by an engineered mutant impaired in quorum-sensing (QS) signalling with the corresponding wild-type led to the conclusion that QS is not involved in the regulation of volatile production in B. ambifaria LMG strain 1918

The aminotransferase Aat initiates 3-phenyllactic acid biosynthesis in Pediococcus acidilactici

The function of the aminotransferase Aat (GenBank Protein WP_159211138) from Pediococcus acidilactici FAM 18098 was studied in vivo. For this purpose, the gene was replaced with an erythromycin resistance gene using the temperature-sensitive Escherichia coli-Pediococcus shuttle plasmid pSET4T_Δaat. The knockout was verified by PCR and genome sequencing. Subsequently, the differences between the metabolism of the knockout and of the wild-type strain were investigated by determining the free amino acids and organic acids in culture supernatants. It was found that the knockout mutant no longer synthesized 3-phenyllactic acid (PLA) and 4-hydroxyphenyllactic acid (HPLA). Additionally, the mutant strain no longer catabolized phenylalanine. Metabolic pathway analysis using the KEGG database indicate that P. acidilactici cannot synthesize α-ketoglutarate that is a predominant amino-group acceptor in many transamination reactions. To study the transfer of the amino group of phenylalanine, the wild-type strain was incubated with [15N] phenylalanine. Mass spectrometry showed that during fermentation, [15N] alanine was formed, indicating that pyruvic acid is an amino group acceptor in P. acidilactici. The present study shows that Aat plays a crucial role in PLA/HPLA biosynthesis and pyruvic acid is an amino acceptor in transamination reactions in P. acidilactici

White lupin leads to increased maize yield through a soil fertility-independent mechanism: a new candidate for fighting Striga hermonthica infestation?

Nitrogen (N)-deficiency and lack of phosphorus (P) availability are major constraints to maize yields in Western Kenya. In a two-season field study in the lake Victoria basin, we tested the capacity of white lupin (Lupinus albus (L.), cv. Ultra), as a nitrogen-fixing crop with a highly efficient P-acquisition capacity, to increase maize yields when used as a companion or cover crop, or as a source of organic matter. Each experiment was performed on three different fields (Vertisols) differing in N/P availability, previous cropping history and in levels of infestation by the parasitic weed Striga hermonthica (Del.) Benth. Our results show that white lupin led to significantly higher yields of maize when used as a cover crop. When lupin was grown as a companion crop, it also slightly enhanced the yield of the co-cultivated maize. When lupin shoots were incorporated to the soil, the positive effect of lupin on maize growth was field-dependent and only occurred in the field most heavily infested with S. hermonthica. Despite the beneficial impact on maize yield, no clear effect of lupin on soil N and P availability or on maize N/P uptake were observed. In contrast, lupin significantly inhibited infestation of maize by S. hermonthica: when lupin was grown together with maize in pots inoculated with S. hermonthica, the emergence of the weed was strongly reduced compared to the pots with maize only. This work opens a new range of questions for further research on white lupin and its potential beneficial impact as a S. hermonthica-inhibiting cro