Insect-pathogenic Photorhabdus luminescens bacteria are widely used in biocontrol strategies against pests. Very little is known about the life of these bacteria in the rhizosphere. Here, we show that P. luminescens can specifically react to and interact with plant roots. Understanding the adaptation of P. luminescens in the rhizosphere is highly important for the biotechnological application of entomopathogenic bacteria and could improve future sustainable pest management in agriculture.

Photorhabdus luminescens are Gram-negative bacteria that live in symbiosis with soil nematodes and are simultaneously highly pathogenic towards insects. The bacteria exist in two phenotypically different forms, designated as primary (1°) and secondary (2°) cells. Yet unknown environmental stimuli as well as global stress conditions induce phenotypic switching of up to 50% of 1° to 2° cells. An important difference between the two phenotypic forms is that 2° cells are unable to live in symbiosis with nematodes and are therefore believed to remain in the soil after a successful infection cycle. In this work, we performed a transcriptomic analysis to highlight and better understand the role of 2° cells and their putative ability to adapt to a life in soil. We could confirm that the major phenotypic differences between the two cell forms are mediated at transcriptional level as the corresponding genes were down-regulated in 2° cells. Furthermore, 2° cells seem to be adapted to another environment as we found several differentially expressed genes involved in the cells9 metabolism, motility and chemotaxis as well as stress resistance, which are either up- or down-regulated in 2° cells. As in contrast to 1° cells 2° cells chemotactically responded to different attractants including plant root exudates there is evidence for the rhizosphere as an alternative environment for the 2° cells. Since P. luminescens is biotechnologically used as bio-insecticide, investigation of a putative interaction of 2° cells with plants is also of great interest for agriculture. Importance The biological function and the fate of P. luminescens 2° cells was unclear. Here we performed comparative transcriptomics of P. luminescens 1° and 2° cultures and found several genes that are up- or down-regulated in 2° cells compared to 1° cells not only coding for known phenotypic differences of the two cell forms. Our results suggest that when 1° cells convert to 2° cells, they drastically change their way of life. Thus, 2° cells could easily adapt to an alternative environment such as a free live in the rhizosphere putatively utilizing plant derived compounds as nutrient source. Since 2° cells are not able to re-associate with the nematodes, an alternative lifestyle in the rhizosphere would be conceivable.

Mycosis fungoides (MF) is the most common entity of Cutaneous T cell lymphomas (CTCL) and is characterized by the presence of clonal malignant T cells in the skin. The role of the skin microbiome for MF development and progression are currently poorly understood. Using shotgun metagenomic profiling, real-time qPCR, and T cell receptor sequencing, we compared lesional and nonlesional skin of 20 MF patients with early and advanced MF. Additionally, we isolated Staphylococcus aureus and other bacteria from MF skin for functional profiling and to study the S. aureus virulence factor spa. We identified a subgroup of MF patients with substantial dysbiosis on MF lesions and concomitant outgrowth of S. aureus on plaque-staged lesions, while the other MF patients had a balanced microbiome on lesional skin. Dysbiosis and S. aureus outgrowth were accompanied by ectopic levels of cutaneous antimicrobial peptides (AMPs), including adaptation of the plaque-derived S. aureus strain. Furthermore, the plaque-derived S. aureus strain showed a reduced susceptibility towards antibiotics and an upregulation of the virulence factor spa, which may activate the NF-κB pathway. Remarkably, patients with dysbiosis on MF lesions had a restricted T cell receptor repertoire and significantly lower event-free survival. Our study highlights the potential for microbiome-modulating treatments targeting S. aureus to prevent MF progression.

The enteric enterobacterium Photorhabdus luminescens is already being used as a bioinsecticide since it is highly pathogenic toward a broad range of insects. However, the bacteria exist in two phenotypically different cell types, called 1° and 2° cells.

Purpose: Phenotypic heterogeneity occurs in many bacterial populations: single cells of the same species display different phenotypes, despite being genetically identical. The Gram-negative entomopathogenic bacterium Photorhabdus luminescens is an excellent example to investigate bacterial phenotypic heterogeneity. Its dualistic life cycle includes a symbiotic stage interacting with entomopathogenic nematodes (EPNs) and a pathogenic stage killing insect larvae. P. luminescens appears in two phenotypically different cell forms: the primary (1°) and the secondary (2°) cell variants. While 1° cells are bioluminescent, pigmented, and produce a huge set of secondary metabolites, 2° cells lack all these phenotypes. The main difference between both phenotypic variants is that only 1° cells can undergo symbiosis with EPNs, a phenotype that is absent from 2° cells. Recent comparative transcriptome analysis revealed that genes mediating 1° cell-specific traits are modulated differently in 2° cells. Although it was previously suggested that heterogeneity in P. luminescens cells cultures is not genetically mediated by, e.g., larger rearrangements in the genome, the genetic similarity of both cell variants has not clearly been demonstrated yet. Methods: Here, we analyzed the genomes of both 1° and 2° cells by genome sequencing of each six single 1° and 2° clones that emerged from a single 1° clone after prolonged growth. Using different bioinformatics tools, the sequence data were analyzed for clustered point mutations or genetic rearrangements with respect to the respective phenotypic variant. Result: We demonstrate that isolated clones of 2° cells that switched from the 1° cell state do not display any noticeable mutation and do not genetically differ from 1° cells. Conclusion: In summary, we show that the phenotypic differences in P. luminescens cell cultures are obviously not caused by mutations or genetic rearrangements in the genome but truly emerge from phenotypic heterogeneity.

In bacteria, group-coordinated behavior such as biofilm formation or virulence are often mediated via cell–cell communication, a process referred to as quorum sensing (QS). The canonical QS system of Gram-negative bacteria uses N-acyl homoserine lactones (AHLs) as communication molecules, which are produced by LuxI-type synthases and sensed by cognate LuxR-type receptors. These receptors act as transcriptional regulators controlling the expression of specific genes. Some bacteria harbor LuxR-type receptors lacking a cognate LuxI-type synthases, designated as LuxR solos. Among many other LuxR solos, the entomopathogenic enteric bacterium Photorhabdus luminescens harbors a SdiA-like LuxR solo containing an AHL signal-binding domain, for which a respective signal molecule and target genes have not been identified yet. Here we performed SPR analysis to demonstrate that SdiA acts as a bidirectional regulator of transcription, tightly controlling its own expression and the adjacent PluDJC_01670 (aidA) gene in P. luminescens, a gene supposed to be involved in the colonization of eukaryotes. Via qPCR we could further determine that in sdiA deletion mutant strains, aidA is upregulated, indicating that SdiA negatively affects expression of aidA. Furthermore, the ΔsdiA deletion mutant exhibited differences in biofilm formation and motility compared with the wild-type. Finally, using nanoDSF analysis we could identify putative binding ability of SdiA towards diverse AHLs, but also to plant-derived signals, modulating the DNA-binding capacity of SdiA, suggesting that this LuxR solo acts as an important player in interkingdom signaling between P. luminescens and plants.