The Periplasmic Oxidoreductase DsbA Is Required for Virulence of the Phytopathogen Dickeya solani

In bacteria, the DsbA oxidoreductase is a crucial factor responsible for the introduction of disulfide bonds to extracytoplasmic proteins, which include important virulence factors. A lack of proper disulfide bonds frequently leads to instability and/or loss of protein function; therefore, improper disulfide bonding may lead to avirulent phenotypes. The importance of the DsbA function in phytopathogens has not been extensively studied yet. Dickeya solani is a bacterium from the Soft Rot Pectobacteriaceae family which is responsible for very high economic losses mainly in potato. In this work, we constructed a D. solani dsbA mutant and demonstrated that a lack of DsbA caused a loss of virulence. The mutant bacteria showed lower activities of secreted virulence determinants and were unable to develop disease symptoms in a potato plant. The SWATH-MS-based proteomic analysis revealed that the dsbA mutation led to multifaceted effects in the D. solani cells, including not only lower levels of secreted virulence factors, but also the induction of stress responses. Finally, the outer membrane barrier seemed to be disturbed by the mutation. Our results clearly demonstrate that the function played by the DsbA oxidoreductase is crucial for D. solani virulence, and a lack of DsbA significantly disturbs cellular physiology.

Nano-Metals Forming Bacteria in Egypt. II. Efficacy towards Some Bacterial Soft Rot/Blackleg Genera ex vivo and Bio-Control in vivo as Eco-Friendly Therapeutic against Dickeya Solani

The antibacterial activity of Cu, Fe, Co, and ZnNPs which were formed by Enterococcus thailandicus, Pseudomonas putida, Marinobacter hydrocarbonoclasticus, and P. geniculate sequentially was tested against some soft rot/blackleg genera. The effects of NPs were recorded on bacterial DNA, proteins, and carbohydrates concentration of Pectobacterium carotovorum, Enterobacter cloacae (soft rot), and Dickeya solani (soft rot/blackleg). Treated cells showed degradation in the DNA isolated, and a decrease in proteins, and carbohydrates concentration compared with untreated cells. The treated cells using SEM showed collapsed, and small pits in the cell wall. Internal changes using TEM showed penetration of NPs inside the tested bacterial cells, the appearance of periplasmic space, formation vacuoles, and condensation of cytoplasm. Disease severity ex vivo of tuber infected with tested genera demonstrated that NPs treatment didn't show any rotted tissue compared with untreated. FeNPs were tested to control soft rot/blackleg disease caused by D. solani in comparison with copper pesticide. Present data recorded an increase in shoot and root length, in addition to, increase in dry and fresh weight, compared with either infected or healthy plants. In studying the ability of treated potato (Solanum tuberosum) seedlings with NPs to uptake and accumulate FeNPs from the soil, ICP-OES recorded a small increase in Fe content of treated plants compared with untreated. FeNPs can be used to control soft rot/blackleg disease caused by D. solani instead of copper pesticide and can be considered as a new and alternative approach to traditional disease management methods, and also increase the nutritional value of the plants

Pathogenicity and Relative Abundance of Dickeya and Pectobacterium Species in Switzerland: An Epidemiological Dichotomy

Pectobacterium and Dickeya species are the causal agents of blackleg and soft rot diseases in potatoes. The main pathogenic species identified so far on potatoes are Dickeya dianthicola, Dickeya solani, Pectobacterium atrosepticum, Pectobacterium brasiliense, Pectobacterium carotovorum, and Pectobacterium parmentieri. Ten years ago, the most prevalent Soft Rot Pectobacteriaceae in Europe were the Dickeya species, P. atrosepticum and P. carotovorum, with some variations among countries. Since then, a drastic increase in the abundance of P. brasiliense has been observed in most European countries. This shift is difficult to explain without comparing the pathogenicity of all Dickeya and Pectobacterium species. The pathogenicity of all the above-mentioned bacterial species was assessed in field trials and in vitro tuber slice trials in Switzerland. Two isolates of each species were inoculated by soaking tubers of cv. Desiree in a suspension of 105 CFU/mL, before planting in the field. For all trials, the Dickeya species were the most virulent ones, but long-term strain surveys performed in Switzerland indicate that P. brasiliense is currently the most frequent species detected. Our results show that the pathogenicity of the species is not the main factor explaining the high prevalence of P. brasiliense and P. parmentieri in the Swiss potato fields.

Dickeya solani D s0432-1 produces an arsenal of secondary metabolites with anti-prokaryotic and anti-eukaryotic activities against bacteria, yeasts, fungi, and aphids.

The necrotrophic plant pathogenic bacterium Dickeya solani is a new invader of potato agrosystem in Europe. All isolated strains of D. solani contain several large polyketide/fatty acid/non-ribosomal peptide synthetase clusters. Analogy with genes described in other bacteria, suggests that two clusters are involved in the production of secondary metabolites of the oocydin and zeamine family. In this study, we constructed by an approach of reverse genetics mutants affected in the three secondary metabolite clusters ssm, ooc and zms in order to compare the phenotype of the D. solani strain D s0432-1 with its derived mutants. We demonstrated that the zeamine cluster inhibits growth of gram-positive and gram-negative bacteria. It is also implicated in a toxicity against aphids. The oocydin cluster inhibits growth of fungi of the phylum Ascomycota. Finally, we unveiled the function of a new secondary metabolite cluster (ssm, for solani secondary metabolite), only conserved in some Dickeya species. This cluster produces a secondary metabolite inhibiting yeasts. D. solani therefore produces several molecules that are toxic to a wide range of living and potentially interacting organisms, from bacteria to insects. The expression of these secondary metabolite pathways could contribute to the rapid spread of D. solani in Europe.

Nano-metals forming bacteria in Egypt. I. Synthesis, characterization and effect on some phytopathogenic bacteria in vitro

Bacterial metal reducers were isolated from water samples collected from harsh condition locations in Egypt. Four selected isolates were identified as Enterococcus thailandicus, Pseudomonas putida, Marinobacter hydrocarbonoclasticus, and P. geniculata for Copper (Cu), Iron (Fe), Cobalt (Co) and Zinc (Zn) Nanoparticles (NPs) production sequentially. Nitrate reductase enzyme was assayed for bacterial isolates which demonstrated that P. putida, and M. hydrocarbonoclasticus have the maximum enzyme production. The produced NPs were characterized by using XRD, TEM, UV–VIS spectroscopy. Magnetic properties for all selected metals NPs were measured using Vibrating Sample Magnetometer (VSM) and demonstrated that FeNPs recorded the highest magnetization value. The antibacterial activity of selected metals NPs was tested against some phytopathogenic bacteria causing the following diseases: soft rot (Pectobacterium carotovorum, Enterobacter cloacae), blackleg (Pectobacterium atrosepticum and Dickeya solani), brown rot (Ralstonia solanacearum), fire blight (Erwinia amylovora) and crown gall (Agrobacterium tumefaciens). All metals NPs showed an antagonistic effect against the tested isolates, particularly, FeNPs showed the highest antibacterial activity followed by CuNPs, and ZnNPs. Due to the small size, high reactivity, and large surface area of biologically synthesized NPs, they are used as a good disinfector, and can be considered as a new and alternative approach to traditional disease management methods.

Early Emergence of Dickeya solani Revealed by Analysis of Dickeya Diversity of Potato Blackleg and Soft Rot Causing Pathogens in Switzerland

Blackleg and soft rot in potato caused by Pectobacterium and Dickeya enterobacteral genera are among the most destructive bacterial diseases in this crop worldwide. In Europe, over the last century, Pectobacterium spp. were the predominant causal agents of these diseases. As for Dickeya, before the large outbreak caused by D. solani in the 2000s, only D. dianthicola was isolated in Europe. The population dynamics of potato blackleg causing soft rot Pectobacteriaceae was, however, different in Switzerland as compared to that in other European countries with a high incidence (60 up to 90%) of Dickeya species (at the time called Erwinia chrysanthemi) already in the 1980s. To pinpoint what may underlie this Swiss peculiarity, we analysed the diversity present in the E. chrysanthemi Agroscope collection gathering potato isolates from 1985 to 2000s. Like elsewhere in Europe during this period, the majority of Swiss isolates belonged to D. dianthicola. However, we also identified a few isolates, such as D. chrysanthemi and D. oryzeae, two species that have not yet been reported in potatoes in Europe. Interestingly, this study allowed the characterisation of two “early” D. solani isolated in the 1990s. Genomic comparison between these early D. solani strains and strains isolated later during the large outbreak in the 2000s in Europe revealed only a few SNP and gene content differences, none of them affecting genes known to be important for virulence.

High-quality complete genome resource of plant pathogenic bacterium Dickeya solani strain IPO 2019 isolated from Hyacinthus orientalis

Dickeya solani is an emerging plant pathogenic bacterium, causing disease symptoms in a variety of agriculturally relevant crop species worldwide. To date a number of D. solani genomes have been sequenced and characterized, the great majority of these genomes have however come from D. solani strains isolated from potato (Solanum tuberosum L.) and not from other plant hosts. Herewith, we present the first complete, high-quality genome of D. solani strain IPO 2019 (LMG 25990) isolated from ornamental plant Hyacinthus orientalis. The genome of D. solani strain IPO 2019 consists of one chromosome of 4,919,542 bp., with a GC content of 56.2% and no plasmids. The genome contains 4502 annotated features, 22 rRNA genes, 73 tRNA genes and 1 CRISPRS. We believe that the information of this high-quality, complete, closed genome of D. solani strain isolated from host plant different than potato (i.e. hyacinth) will provide resources for comparative genomic studies as well as for analyses targeting adaptation and ecological fitness mechanisms present in Dickeya solani species.

Dickeya solani (black leg disease of potato).

Dickeya solani is a bacterial pathogen of potato thought to have crossed from horticultural crops in Northern Europe in 2005-2006. The earliest reports of disease on potato are from Belgium and the Netherlands, though it is now present in most European countries and Israel. Symptoms range from blackleg to top wilt in the growing plant and soft rotting of tubers. Symptoms are indistinguishable from those caused by Pectobacterium atrosepticum, P. carotovorum, P. parmentieri (formerly P. wasabiae) (Khayi et al., 2016) and D. dianthicola and control is usually reliant on seed certification schemes to mitigate its worst effects. It should be noted that most losses are attributable to the certification process itself though losses as high as 30% have been recorded in crops grown in Israel. There is no evidence of varietal resistance in potato. D. solani is a highly clonal organism highlighting its recent emergence as a pathogen but also the vulnerability of Europe's highly integrated potato production system.

Co-occurrence of latent Dickeya and Pectobacterium species in potato seed tuber samples from northern Finland

Recent methodological developments have uncovered the etiological diversity of the potato blackleg and soft rot Pectobacteriaceae. At least five species in the genera Dickeya and Pectobacterium have been confirmed to cause blackleg on potatoes in Finland. The bacteria are seed borne and remain latent in the tuber until conditions favourable for growth, multiplication and infection prevail. Tubers could be infected by one or more of these species. This short communication is based on the results of molecular detection data collected for more than 14 years from potato seed lots produced in Finland. Diagnostic PCR assay specific to Dickeya solani, Pectobacterium atrosepticum, Pectobacterium carotovorum, P. brasiliense and P. parmentieri revealed that potatoes are infected by one or more of these species; it also revealed that single species infection is more common than multiple colonization. An event of simultaneous occurrences of different strains from the Pectobacterium species appears to be more frequent than that observed between Dickeya and Pectobacterium species. The absence of co-occurrence of Dickeya solani and Pectobacterium atrosepticum is intriguing.