The host specificity of Fusarium oxysporum (Fox) formae speciales has been reported to be linked to effector proteins known as Secreted in Xylem (SIX). These genes are associated with the non-autonomous mobile element miniature impala (mimp), normally distributed on the accessory chromosomes. The pattern of mimp associated with effector genes has been used to predict candidate effector profiles which characterize Fox formae speciales. In this study, we demonstrate the pathogenicity of strains Fusarium oxysporum f.sp. radicis-lycopersici (Forl) ZUM2407 and Fusarium oxysporum f.sp. radicis-cucumerinum (Forc) V03-2g in a common host plant (cucumber) and compare their genomes. The Forl ZUM2407 genome lacks SIX genes and their homologs, in contrast to Forc V03-2g. We predicted the total number of mimp elements in the genome of Forl ZUM2407 to be three-fold less than that of Forc V03-2g (10 and 36 copies, respectively). The mimp distribution pattern in Forl ZUM2407 was completely different from that present in Forc V03-2g. Candidate effector profile analysis did not predict that Forl ZUM2407 was able to infect cucumber plants like Forc V03-2g. Therefore, we assume that Forl ZUM2407 has a different type of genome organization associated with pathogenicity, whose effector profile cannot be described using the mimp-based approach.

Bacillus species have gained much attention based on their phenotypic characteristics and their genetic architecture as biological control agents and plant growth-promotor with bioremediation potential. In this study, we analyzed the whole genome of a novel strain, Bacillus glycinifermentans MGMM1, isolated from the rhizosphere of a weed plant (Senna occidentalis) and assayed its phenotypic characteristics, as well as antifungal and biocontrol ability. The whole genome analysis of MGMM1 identified 4259 putative coding sequences, with an encoding density of 95.75% attributed to biological functions, including genes involved in stimulating plant growth, such as acetolactate synthase, alsS, and genes involved in the resistance to heavy metal antimony (arsB and arsC). AntiSMASH revealed the presence of biosynthetic gene clusters plipastatin, fengycin, laterocidine, geobacillin II, lichenysin, butirosin A and schizokinen. Tests in vitro confirmed that MGMM1 exhibited antifungal activity against Fusarium oxysporum f.sp. radicis-lycopersici (Forl) ZUM2407, Alternaria alternata, F. graminearum and F. spp. and produce protease, lipase amylase and cellulase. Bacillus glycinifermentans MGMM1 demonstrated proteolytic (4.82 ± 1.04 U/mL), amylolytic (0.84 ± 0.05 U/mL) and cellulosic (0.35 ± 0.02 U/mL) enzymatic activities, as well as indole-3-acetic acid production (48.96 ± 1.43 μg/mL). Moreover, the probiotic strain MGMM1 demonstrated a high biocontrol potential of inhibiting (up to 51.45 ± 8.08%) the development of tomato disease caused by Forl ZUM2407. These results suggest that B. glycinifermentans MGMM1 has significant potential as a biocontrol, plant growth-promoting agent in agriculture.

Fusarium oxysporum f. sp. radicis-cucumerinum (Forc) V03-2g and Fusarium oxysporum f. sp. radicis-lycopersici (Forl) ZUM2407 both cause foot and root rot in cucumber, but differ in host range. Forc V03-2g possesses Secreted in Xylem (SIX) effector genes, whereas Forl ZUM2407 does not, raising questions about their distinct infection strategies on this host. Using comparative transcriptomic analysis (in cucumber at 7 and 14 days post-inoculation (dpi) and in tomato at 2 dpi) we show that Forl ZUM2407 induces a delayed defense response in cucumber compared to Forc V03-2g. In turn, Forc V03-2g rapidly activates accessory chromosome effectors on cucumber, while Forl ZUM2407 initially deploys core chromosome genes, activating distinct from Forc V03-2g accessory genes only by 14 dpi. Thereby, Forc V03-2g and Forl ZUM2407 use distinct accessory gene repertoires (unique to each strain) and distinct core gene transcription strategies to infect the same host.

Multi-strain consortia are expected to protect plants more reliably than single strains, yet this is rarely tested directly, and the behavior of introduced strains in the field is difficult to follow. We assembled a consortium of Stenotrophomonas rhizophila MGMM118, Bacillus halotolerans MGMM119, Pseudomonas grimontii MGMM120 and Pseudomonas viciae MGMM121 strains isolated from the winter wheat rhizosphere and characterized their hydrolytic activities, auxin production, and analyzed their genomes. The consortium increased shoot and root length of wheat. In biocontrol experiments on barley, the consortium reduced the root rot disease index caused by Fusarium culmorum by approximately ten-fold (5.6% vs. 59.7%), outperforming each individual strain (11.9–17.9%). The consortium also reduced Fusarium root rot in tomato by two-fold. In a field experiment on wheat, strain-specific qPCR detected all introduced microorganisms in the wheat rhizosphere at up to 106 CFU per gram of soil, demonstrating different dynamics for each individual strain. Metabarcoding showed that community structure depended on plant developmental stage rather than on the consortium application. Since metabarcoding could not reliably identify the introduced strains, we suppose that strain-specific qPCR is a more appropriate tool for monitoring of inoculants, particularly in complex soil microbial communities.

Microorganisms are fundamental drivers of soil productivity, mediating nutrient cycling and pathogen suppression. In this study, we evaluated changes in the fungal community in the soil of barley (Hordeum vulgare L.) in a field experiment involving the application of a consortium of Paenibacillus pabuli, Priestia megaterium, Pseudomonas koreensis, and Pseudomonas orientalis. Seed pretreatment and seed pretreatment followed by rhizosphere drenching at different growth stages were implemented. Regarding fungal communities in bulk soil, the rhizospheres of untreated and treated plants were characterized based on full-length ribosomal RNA gene (18S-5.8S-28S) metabarcoding sequencing. Despite the compositional shifts, no statistical differences were observed among the alpha diversity metrics. Seed treatment resulted in long-term, targeted suppression of Fusarium graminearum, Fusarium fujikuroi, Fusarium musae, and Fusarium verticillioides from the booting through flowering and dough development stages, outperforming seed pretreatment followed by rhizosphere drenching. A low-modularity network was observed in the rhizosphere of untreated plants. Seed treatment fostered a highly interconnected and uniform network with low hub-betweenness scores. Rhizosphere drenching of pretreated seeds shifted the network topology toward higher hub-betweenness scores, reducing their connectivity by up to 10% in the rhizosphere and bulk soil. These findings provide a framework for optimizing the soil ecosystem for sustainable agriculture.