Draft Genome Sequence of Pseudomonas brassicacearum Strain UTMN3, a Biological Control Agent from the Rhizosphere of Pisum sativum

Here, we announce and describe the draft genome sequence of Pseudomonas brassicacearum UTMN3, which contains 40 contigs comprising 6,658,810 bp, with a GC content of 60.9%. The genome contains 5,825 protein-coding genes and 65 RNA-coding genes. The genome of UTMN3 contains several genes that are likely contributors to plant protection.

Increasing the antimicrobial activity of Pseudomonas brassicacearum S-1 by inactivation of lysR3 and mtfA genes

Site-specific markerless inactivation of genes lysR3 and mtfA of bacteria Pseudomonas brassicacearum S-1 was carried out. It was shown that strains P. brassicacearum S-1-lysR3, P. brassicacearum S-1-mtfA and P. brassicacearum S-1-lysR3-mtfA demonstrate increased antagonistic activity against a number of phytopathogens 1.2–1.6 times higher compared with the original strain. In the laboratory experiment, mutant variants did not affect growth and development of tomato, cucumber and parsley plants. Treating tomato and cucumber seeds with a solution of the culture of the strain P. brassicacearum S-1-lysR3-mtfA led to the increase of seedlings roots length by 1.4 times compared with the original strain. We also determined that the examined strains were able to survive in the soil microbial consortium.

Evaluation of the phytotoxicity of 2,4-diacetylphloroglucinol and Pseudomonas brassicacearum Q8r1-96 on different wheat cultivars

Pseudomonas brassicacearum Q8r1-96 and other 2,4-diacetylphloroglucinol (DAPG)-producing pseudomonads of the Pseudomonas fluorescens complex possess both biocontrol and growth-promoting properties and play an important role in suppression of take-all of wheat in the Pacific Northwest (PNW) of the United States. However, P. brassicacearum can also reduce seed germination and cause root necrosis on some wheat cultivars. We evaluated the effect of Q8r1-96 and DAPG on the germination of 69 wheat cultivars that have been or currently are grown in the PNW. Cultivars varied widely in their ability to tolerate P. brassicacearum or DAPG. The frequency of germination of the cultivars ranged from 0 to 0.87 and from 0.47 to 0.90 when treated with Q8r1-96 and DAPG, respectively. There was a significant positive correlation between the frequency of germination of cultivars treated with Q8r1-96 in assays conducted in vitro and in the greenhouse. The correlation was greater for spring than for winter cultivars. In contrast, the effect of Q8r1-96 on seed germination was not correlated with that of DAPG alone, suggesting that DAPG is not the only factor responsible for the phytotoxicity of Q8r1-96. Three wheat cultivars with the greatest tolerance and three cultivars with the least tolerance to Q8r1-96 were tested for their ability to support root colonization by strain Q8r1-96. Cultivars with the greatest tolerance supported significantly greater populations of strain Q8r1-96 than those with the least tolerance to the bacteria. Our results show that wheat cultivars differ widely in their interaction with P. brassicacearum and the biocontrol antibiotic DAPG.

GENOME ANALYSIS OF Pseudomonas brassicacearum S-1 – AN ANTAGONIST OF CROP PATHOGENS

The strain Pseudomonas brassicacearum S-1 is the basis of the biopesticide “Ecogreen”, which is used to control pathogens infecting vegetable and green spicy crops in small-scale hydroponics. Aim. The purpose of this work was to sequence and analyze the nucleotide sequence of the genome of strain P. brassicacearum S-1 (GenBank accession number CP045701). Methods. Whole-genome sequencing was performed by both MiSeq (Illuminа) and MinION (Oxford Nanopore). Analysis of the genome sequence was performed with a number of bioinformatics programs. Results. The genome of the P. brassicacearum S-1 strain comprising a single circular 6 577 561-bp chromosome with GC content of 60.8 %. Genome analysis revealed genes that constitute valuable biotechnological potential of the S-1 strain and determine synthesis of a wide range of secondary metabolites. Moreover, mobile genetic elements, prophages and short repetitive sequences were identified in the S-1 genome. Conclusions. Detected genetic determinants, which are responsible for the synthesis of practically valuable compounds, indicate a significant potential of the P. brassicacearum S-1 strain as a biocontrol agent.

Selection and characterization of bacteria – the basis of microbial preparation improving quality of lawns

During the study two bacterial strains Pseudomonas brassicacearum S-1 and Flavobacterium johnsoniae Ч1 were selected to be further used as the basis of microbial preparation to improve the quality of lawns.

Inactivation of genes lysR and mtfA increases antagonistic activity of bacteria Pseudomonas brassicacearum S-1

Genes lysR and mtfA localized in chromosome of bacteria P. brassicacearum S 1 were inactivated. The role of these genes in the expression of strain antagonistic activity against several bacterial and fungal pathogens was investigated. The technique of markerless mutagenesis of bacteria P. brassicacearum was optimized.

Selection of the Root Endophyte Pseudomonas brassicacearum CDVBN10 as Plant Growth Promoter for Brassica napus L. Crops

Rapeseed (Brassica napus L.) is an important crop worldwide, due to its multiple uses, such as a human food, animal feed and a bioenergetic crop. Traditionally, its cultivation is based on the use of chemical fertilizers, known to lead to several negative effects on human health and the environment. Plant growth-promoting bacteria may be used to reduce the need for chemical fertilizers, but efficient bacteria in controlled conditions frequently fail when applied to the fields. Bacterial endophytes, protected from the rhizospheric competitors and extreme environmental conditions, could overcome those problems and successfully promote the crops under field conditions. Here, we present a screening process among rapeseed bacterial endophytes to search for an efficient bacterial strain, which could be developed as an inoculant to biofertilize rapeseed crops. Based on in vitro, in planta, and in silico tests, we selected the strain Pseudomonas brassicacearum CDVBN10 as a promising candidate; this strain produces siderophores, solubilizes P, synthesizes cellulose and promotes plant height in 5 and 15 days-post-inoculation seedlings. The inoculation of strain CDVBN10 in a field trial with no addition of fertilizers showed significant improvements in pod numbers, pod dry weight and shoot dry weight. In addition, metagenome analysis of root endophytic bacterial communities of plants from this field trial indicated no alteration of the plant root bacterial microbiome; considering that the root microbiome plays an important role in plant fitness and development, we suggest this maintenance of the plant and its bacterial microbiome homeostasis as a positive result. Thus, Pseudomonas brassicacearum CDVBN10 seems to be a good biofertilizer to improve canola crops with no addition of chemical fertilizers; this the first study in which a plant growth-promoting (PGP) inoculant specifically designed for rapeseed crops significantly improves this crop’s yields in field conditions.

Exploring the Pathogenicity of Pseudomonas brassicacearum Q8r1-96 and Other Strains of the Pseudomonas fluorescens Complex on Tomato

Pseudomonas brassicacearum and related species of the P. fluorescens complex have long been studied as biocontrol and growth-promoting rhizobacteria involved in suppression of soilborne pathogens. We report here that P. brassicacearum Q8r1-96 and other 2,4-diacetylphloroglucinol (DAPG)-producing fluorescent pseudomonads involved in take-all decline of wheat in the Pacific Northwest of the United States can also be pathogenic to other plant hosts. Strain Q8r1-96 caused necrosis when injected into tomato stems and immature tomato fruits, either attached or removed from the plant, but lesion development was dose dependent, with a minimum of 106 CFU ml−1 required to cause visible tissue damage. We explored the relative contribution of several known plant-microbe interaction traits to the pathogenicity of strain Q8r1-96. Type III secretion system (T3SS) mutants of Q8r1-96, injected at a concentration of 108 CFU ml−1, were significantly less virulent, but not consistently, as compared with the wild-type strain. However, a DAPG-deficient phlD mutant of Q8r1-96 was significantly and consistently less virulent as compared with the wild type. Strain Q8r1-96acc, engineered to over express ACC deaminase, caused a similar amount of necrosis as the wild type. Cell-free culture filtrates of strain Q8r1-96 and pure DAPG also cause necrosis in tomato fruits. Our results suggest that DAPG plays a significant role in the ability of Q8r1-96 to cause necrosis of tomato tissue, but other factors also contribute to the pathogenic properties of this organism.

Friend or foe? Exploring the fine line between Pseudomonas brassicacearum and phytopathogens

Pseudomonas brassicacearum is one of over fifty species of bacteria classified into the P. fluorescens group. Generally considered a harmless commensal, these bacteria are studied for their plant-growth promotion (PGP) and biocontrol characteristics. Intriguingly, P. brassicacearum is closely related to P. corrugata , which is classified as an opportunistic phytopathogen. Twenty-one P. brassicacearum genomes have been sequenced to date. In the current review, genomes of P. brassicacearum and strains from the P. corrugata clade were mined for regions associated with PGP, biocontrol and pathogenicity. We discovered that ‘beneficial’ bacteria and those classified as plant pathogens have many genes in common; thus, only a fine line separates beneficial/harmless commensals from those capable of causing disease in plants. The genotype and physiological state of the plant, the presence of biotic/abiotic stressors, and the ability of bacteria to manipulate the plant immune system collectively contribute to how the bacterial-plant interaction plays out. Because production of extracellular metabolites is energetically costly, these compounds are expected to impart a fitness advantage to the producer. P. brassicacearum is able to reduce the threat of nematode predation through release of metabolites involved in biocontrol. Moreover this bacterium has the unique ability to form biofilms on the head of Caenorhabditis elegans, as a second mechanism of predator avoidance. Rhizobacteria, plants, fungi, and microfaunal predators have occupied a shared niche for millions of years and, in many ways, they function as a single organism. Accordingly, it is essential that we appreciate the dynamic interplay among these members of the community.