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Priming plants with beneficial microbes can establish rapid and robust resistance against numerous pathogens. Here, compelling evidence is provided that the treatment of rapeseed plants with Trichoderma harzianum OMG16 and Bacillus velezensis FZB42 induces defence activation against Verticillium longisporum infection. The relative expressions of the JA biosynthesis genes LOX2 and OPR3, the ET biosynthesis genes ACS2 and ACO4 and the SA biosynthesis and signalling genes ICS1 and PR1 were analysed separately in leaf, stem and root tissues using qRT-PCR. To successfully colonize rapeseed roots, the V. longisporum strain 43 pathogen suppressed the biosynthesis of JA, ET and SA hormones in non-primed plants. Priming led to fast and strong systemic responses of JA, ET and SA biosynthesis and signalling gene expression in each leaf, stem and root tissue. Moreover, the quantification of plant hormones via UHPLC-MS analysis revealed a 1.7- and 2.6-fold increase in endogenous JA and SA in shoots of primed plants, respectively. In roots, endogenous JA and SA levels increased up to 3.9- and 2.3-fold in Vl43-infected primed plants compared to non-primed plants, respectively. Taken together, these data indicate that microbial priming stimulates rapeseed defence responses against Verticillium infection and presumably transduces defence signals from the root to the upper parts of the plant via phytohormone signalling. [ABSTRACT FROM AUTHOR]

The persistence of beneficial microorganisms in the rhizosphere or surrounding soil following their application is a prerequisite for the effective interaction with the plant or indigenous microbial communities in the respective habitats. The goal of the study was to analyze the establishment and persistence of the applied beneficial Trichoderma harzianum (OMG16) strain in the maize root-associated soil depending on agricultural practice (soil management practice, N-fertilizer intensity) in a field experiment. A rapid identification of the inoculated strain OMG16 is essential for its monitoring. We used a culture-based approach coupled to matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis for the rapid identification of the inoculated Trichoderma strain as part of the beneficial microbe consortium (BMc). We isolated 428 fungal isolates from eight treatments of the field experiment. Forty eight percent of the isolated fungi equivalent to 205 fungal isolates were identified as Trichoderma, of which 87% (=179 isolates) were obtained from the fields inoculated with BMc. Gene sequence analysis showed a high similarity of the MALDI-TOF MS-identified Trichoderma, with that of the inoculated Trichoderma harzianum OMG16 confirming the re-isolation of the added beneficial fungus. This study highlighted the use of MALDI-TOF MS analysis as a quick, cost-effective detection and efficient monitoring tool for microbial-based bioinoculants in the field. [ABSTRACT FROM AUTHOR]

The long-term effects of agricultural management such as different fertilization strategies on soil microbiota and soil suppressiveness against plant pathogens are crucial. Therefore, the suppressiveness of soils differing in fertilization history was assessed using two Rhizoctonia solani isolates and their respective host plants (lettuce, sugar beet) in pot experiments. Further, the effects of fertilization history and the pathogen R. solani AG1-IB on the bulk soil, root-associated soil and rhizosphere microbiota of lettuce were analyzed based on amplicon sequencing of the 16S rRNA gene and ITS2 region. Organic fertilization history supported the spread of the soil-borne pathogens compared to long-term mineral fertilization. The fertilization strategy affected bacterial and fungal community composition in the root-associated soil and rhizosphere, respectively, but only the fungal community shifted in response to the inoculated pathogen. The potential plant-beneficial genus Talaromyces was enriched in the rhizosphere by organic fertilization and presence of the pathogen. Moreover, increased expression levels of defense-related genes in shoots of lettuce were observed in the soil with organic fertilization history, both in the absence and presence of the pathogen. This may reflect the enrichment of potential plant-beneficial microorganisms in the rhizosphere, but also pathogen infestation. However, enhanced defense responses resulted in retarded plant growth in the presence of R. solani (plant growth/defense tradeoff). [ABSTRACT FROM AUTHOR]

Using biological fertilizers and pesticides based on beneficial soil microbes in order to reduce mineral fertilizers and chemical pesticides in conventional agriculture is still a matter of debate. In this regard, a European research project seeks to elucidate the role of root-endophytic fungi and to develop molecular tools to trace and quantify these fungi in the rhizosphere and root tissue. To do this, the draft genome sequence of the biocontrol fungus Trichoderma virens (T. virens) was screened for simple sequence repeats (SSRs) and primers were developed for 12 distinct loci. Primers were evaluated using a global collection of ten isolates where an average of 7.42 alleles per locus was detected. Nei's standard genetic distance ranged from 0.18 to 0.27 among the isolates, and the grand mean of haploid diversity in AMOVA analysis was 0.693 ± 0.019. Roots of tomato plants were inoculated with different strains and harvested six weeks later. Subsequent PCR amplification identified root-endophytic strains and co-colonization of roots by different strains. Markers were applied to qPCR to quantify T. virens strains in root tissue and to determine their identity using allele-specific melting curve analysis. Thus, the root-endophytic lifestyle of T. virens was confirmed, strains in roots were quantified and simultaneous colonization of roots by different strains was observed. [ABSTRACT FROM AUTHOR]

The use of plant growth-promoting microorganisms (PGPMs) as bio-effectors (BEs) to improve the nutrient acquisition of crops has a long history. However, limited reproducibility of the expected effects still remains a major challenge for practical applications. Based on the hypothesis that the expression of PGPM effects depends on soil type and the properties of the applied fertilizers, in this study, the performance of selected microbial inoculants was investigated for two contrasting low-fertility soils supplied with different organic and inorganic fertilizers. Greenhouse experiments were conducted with tomato on an alkaline sandy loam of pH 7.8 and an acidic loamy sand of pH 5.6 with limited phosphate (P) availability. Municipal waste compost, with and without poultry manure (PM), rock phosphate (RP), stabilized ammonium, and mineral nitrogen, phosphorus and potassium (NPK) fertilization were tested as fertilizer variants. Selected strains of Bacillus amyloliquefaciens (Priest et al. 1987) Borriss et al. 2011 (FZB42) and Trichoderma harzianum Rifai (OMG16) with proven plant growth-promoting potential were used as inoculants. On both soils, P was identified as a major limiting nutrient. Microbial inoculation selectively increased the P utilization in the PM-compost variants by 116% and 56% on the alkaline and acidic soil, while RP utilization was increased by 24%. This was associated with significantly increased shoot biomass production by 37–42%. Plant growth promotion coincided with a corresponding stimulation of root growth, suggesting improved spatial acquisition of soluble soil P fractions, associated also with improved acquisition of nitrogen (N), potassium (K), magnesium (Mg), and calcium (Ca). There was no indication for mobilization of sparingly soluble Ca phosphates via rhizosphere acidification on the alkaline soil, and only mineral NPK fertilization reached a sufficient P status and maximum biomass production. However, on the moderately acidic soil, FZB42 significantly stimulated plant growth of the variants supplied with Ca–P in the form of RP + stabilized ammonium and PM compost, which was equivalent to NPK fertilization; however, the P nutritional status was sufficient only in the RP and NPK variants. The results suggest that successful application of microbial biofertilizers requires more targeted application strategies, considering the soil properties and compatible fertilizer combinations. [ABSTRACT FROM AUTHOR]