Characterization of Novel Bacteriophages for Biocontrol of Bacterial Blight in Leek Caused by Pseudomonas syringae pv. porri

Rombouts, Sigrid; Volckaert, Anneleen; Venneman, Sofie; Declercq, Bart; Vandenheuvel, Dieter; Allonsius, Camille Nina; Malderghem, Cinzia Van; Jang, Ho Bin; Briers, Yves; Noben, Jean-Paul; Klumpp, Jochen; Vaerenbergh, Johan Van; Maes, Martine; Lavigne, Rob

doi:10.3389/fmicb.2016.00279

Cited by 90 publications

(93 citation statements)

References 64 publications

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“…Lytic bacteriophages have shown promise in protecting tomato plants from wilting caused by Ralstonia solanacearum (17,18), protecting oranges from citrus canker caused by Xanthomonas axonopodis (19), protecting leeks from bacterial blight caused by Pseudomonas syringae pv. porri (20), and protecting kiwi from canker caused by Pseudomonas syringae pv. actinidiae (21).…”

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confidence: 99%

Expression of a Peptidoglycan Hydrolase from Lytic Bacteriophages Atu_ph02 and Atu_ph03 Triggers Lysis of Agrobacterium tumefaciens

Attai

Rimbey

Smith

et al. 2017

Appl Environ Microbiol

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To provide food security, innovative approaches to preventing plant disease are currently being explored. Here, we demonstrate that lytic bacteriophages and phage lysis proteins are effective at triggering lysis of the phytopathogen Agrobacterium tumefaciens. Phages Atu_ph02 and Atu_ph03 were isolated from wastewater and induced lysis of C58-derived strains of A. tumefaciens. The coinoculation of A. tumefaciens with phages on potato discs limited tumor formation. The genomes of Atu_ph02 and Atu_ph03 are nearly identical and are ϳ42% identical to those of T7 supercluster phages. In silico attempts to find a canonical lysis cassette were unsuccessful; however, we found a putative phage peptidoglycan hydrolase (PPH), which contains a C-terminal transmembrane domain. Remarkably, the endogenous expression of pph in the absence of additional phage genes causes a block in cell division and subsequent lysis of A. tumefaciens cells. When the presumed active site of the N-acetylmuramidase domain carries an inactivating mutation, PPH expression causes extensive cell branching due to a block in cell division but does not trigger rapid cell lysis. In contrast, the mutation of positively charged residues at the extreme C terminus of PPH causes more rapid cell lysis. Together, these results suggest that PPH causes a block in cell division and triggers cell lysis through two distinct activities. Finally, the potent killing activity of this single lysis protein can be modulated, suggesting that it could be engineered to be an effective enzybiotic. IMPORTANCEThe characterization of bacteriophages such as Atu_ph02 and Atu_ph03, which infect plant pathogens such as Agrobacterium tumefaciens, may be the basis of new biocontrol strategies. First, cocktails of diverse bacteriophages could be used as a preventative measure to limit plant diseases caused by bacteria; a bacterial pathogen is unlikely to simultaneously develop resistances to multiple bacteriophage species. The specificity of bacteriophage treatment for the host is an asset in complex communities, such as in orchards where it would be detrimental to harm the symbiotic bacteria in the environment. Second, bacteriophages are potential sources of enzymes that efficiently lyse bacterial cells. These phage proteins may have a broad specificity, but since proteins do not replicate as phages do, their effect is highly localized, providing an alternative to traditional antibiotic treatments. Thus, studies of lytic bacteriophages that infect A. tumefaciens may provide insights for designing preventative strategies against bacterial pathogens.

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confidence: 99%

Expression of a Peptidoglycan Hydrolase from Lytic Bacteriophages Atu_ph02 and Atu_ph03 Triggers Lysis of Agrobacterium tumefaciens

Attai

Rimbey

Smith

et al. 2017

Appl Environ Microbiol

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“…In Europe, for instance, the registration exists of two phases: registration of the active ingredient at European level by the European Food Safety Authority (EFSA) and next the authorization of the formulated product at member state level leading to bureaucratic difficulties [17]. As phages are highly specific and can only infect a few strains of one bacterial species, they are often combined in a cocktail [25]. In terms of registrations, this means that every phage in the cocktail should be registered as an active ingredient (Regulation (EC) No 1107/2009) and reformulations of the product require reregistrations [4].…”

Section: Discussionmentioning

confidence: 99%

Protection of phage applications in crop production: a patent landscape

Holtappels

Lavigne

Huys

et al. 2019

Preprint

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In agriculture, the prevention and treatment of bacterial infections represents an 10 increasing challenge. Traditional (chemical) methods have been restricted to ensure public health 11 and limit the occurrence of resistant strains. Bacteriophages could be a sustainable alternative. A 12 major hurdle towards the commercial implementation of phage-based biocontrol strategies 13 concerns aspects of regulation and intellectual property protection. Within this study, two datasets 14 have been composed to analyze both scientific publications and patent documents and to get an 15 idea on the focus of R&D by means of an abstract and claim analysis. 137 papers and 49 patent 16 families were found searching public databases, their numbers increasing over time. Within this 17 dataset, the majority of the patent documents were filed by non-profit organizations in Asia. There 18 seems to be a good correlation between the papers and patent documents in terms of targeted 19 bacterial genera. Furthermore, granted patents seem to claim rather broad and cover methods of 20 treatment. This review shows that there is indeed a growing publishing and patenting activity 21 concerning phage biocontrol. Targeted research is needed to further stimulate the exploration of 22 phages within integrated pest management strategies and to deal with bacterial infections in crop 23 production. 24 25 26 29 the first applications of phages focused on human medicine [3], other fields including agriculture 30 soon began to explore the potential of bacteriophages as biocontrol agents [4]. The first isolation of 31 phages infecting plant pathogenic bacteria (PPB) dates back to 1924, when it was shown that 32 Xanthomonas campestris pv. campestris, causing black rot in Brassicaceae, could be lysed by the filtrate 33 of diseased cabbages [4,5]. The following years, interest in phages as biocontrol agents remained 34 relatively high [4]. However, the discovery of broad-spectrum antibiotics and other bactericidal 35 chemicals resulted in a dwindling interest in phage therapy research in general [6].36 Within the agricultural sector, the prevention and treatment of bacterial infections represents an 37 increasing challenge. For farmers, devastating losses by bacterial pathogens are generally estimated 38 to reach 10% of the total production [7] and for some bacterial species like Xanthomonas campestris pv.39 campestris crop yield can be reduced by 25% [8]. Other major threats include Ralstonia solanacearum, 40 Xylella fastidiosa and Pseudomonas syringae pathovars [9]. In recent years, general antibiotics like 41 streptomycin as well as copper-based chemicals have been restricted in crop production to ensure 42 public health and limit the occurrence of resistant strains [10-13]. Because of these restrictions, the 43 search for sustainable, natural biocontrol of PPBs has come to a critical stage, especially considering 44 the increased food production needs [14]. Governments have decided to implement Integrated Pest 45 Management strategies (IPMs) as the stan...

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“…The phage cocktail was able to infect all of the bacterial strains. However, the therapy did not fully prevent or treat infection in the leeks [25]. The investigators of this study used a single application of the phage cocktail and hypothesized that a second application of the cocktail may produce better results [25].…”

Section: Phage Therapy In Plantsmentioning

confidence: 99%

“…However, the therapy did not fully prevent or treat infection in the leeks [25]. The investigators of this study used a single application of the phage cocktail and hypothesized that a second application of the cocktail may produce better results [25]. These investigations are important in understanding the potential uses of phage therapy in the agricultural setting.…”

Section: Phage Therapy In Plantsmentioning

confidence: 99%

A Review of Phage Therapy against Bacterial Pathogens of Aquatic and Terrestrial Organisms

Doss

Culbertson

Hahn

et al. 2017

Viruses

213

183

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Since the discovery of bacteriophage in the early 1900s, there have been numerous attempts to exploit their innate ability to kill bacteria. The purpose of this report is to review current findings and new developments in phage therapy with an emphasis on bacterial diseases of marine organisms, humans, and plants. The body of evidence includes data from studies investigating bacteriophage in marine and land environments as modern antimicrobial agents against harmful bacteria. The goal of this paper is to present an overview of the topic of phage therapy, the use of phage-derived protein therapy, and the hosts that bacteriophage are currently being used against, with an emphasis on the uses of bacteriophage against marine, human, animal and plant pathogens.

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Characterization of Novel Bacteriophages for Biocontrol of Bacterial Blight in Leek Caused by Pseudomonas syringae pv. porri

Cited by 90 publications

References 64 publications

Expression of a Peptidoglycan Hydrolase from Lytic Bacteriophages Atu_ph02 and Atu_ph03 Triggers Lysis of Agrobacterium tumefaciens

Expression of a Peptidoglycan Hydrolase from Lytic Bacteriophages Atu_ph02 and Atu_ph03 Triggers Lysis of Agrobacterium tumefaciens

Protection of phage applications in crop production: a patent landscape

A Review of Phage Therapy against Bacterial Pathogens of Aquatic and Terrestrial Organisms

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