Isolation, characterization and genomic analysis of a novel lytic bacteriophage infecting Agrobacterium tumefaciens

Agrobacterium tumefaciens causes crown gall, and economic losses in important crops, including apple, pear, peach, and almond. Difficulties controlling this disease with conventional pesticides require alternative antibacterial agents. A novel lytic bacteriophage, Agrobacterium phage PAT1 (PAT1), wi...

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Main Authors: Miloud SABRI, Kaoutar EL HANDI, Orges CARA, Angelo DE STRADIS, Toufic ELBEAINO
Format: Article
Language:English
Published: Firenze University Press 2024-11-01
Series:Phytopathologia Mediterranea
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Online Access:https://oajournals.fupress.net/index.php/pm/article/view/15623
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author Miloud SABRI
Kaoutar EL HANDI
Orges CARA
Angelo DE STRADIS
Toufic ELBEAINO
author_facet Miloud SABRI
Kaoutar EL HANDI
Orges CARA
Angelo DE STRADIS
Toufic ELBEAINO
author_sort Miloud SABRI
collection DOAJ
description Agrobacterium tumefaciens causes crown gall, and economic losses in important crops, including apple, pear, peach, and almond. Difficulties controlling this disease with conventional pesticides require alternative antibacterial agents. A novel lytic bacteriophage, Agrobacterium phage PAT1 (PAT1), with high lysis potential against A. tumefaciens, was isolated from wastewater. Interaction between PAT1 and A. tumefaciens cells was investigated using transmission electron microscopy. PAT1 adsorbed, infected, and replicated on A. tumefaciens in ≤30 min. Turbidity assays showed that PAT1 [Multiplicity of Infection (MOI) = 1] inhibited A. tumefaciens growth by 82% for 48 hours. PAT1 was resistant to broad ranges of pH (4 to 10) and temperatures (4 to 60°C). Bioinformatics analyses of the PAT1 genomic sequence showed that the bacteriophage was closely related to Atuphduovirus (Autographiviridae) phages. The PAT1 genome size was 45,040 base pairs with a G+C content of 54.5%, consisting of 54 coding sequences (CDS), of which the functions of 23 CDS were predicted, including an endolysin gene which could be used as an antimicrobial against A. tumefaciens. No lysogenic mediated genes or genes encoding virulence factors, antibiotic resistance, or toxins were detected in PAT1 genome. The bacteriophage showed potential as a biocontrol agent against A. tumefaciens infections, expanding the limited catalogue of lytic A. tumefaciens phages, although efficacy for control of crown gall in planta remains to be evaluated.
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publishDate 2024-11-01
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spelling doaj-art-89c48a27fa9346a3b10bb13f49bb891a2025-08-20T03:08:35ZengFirenze University PressPhytopathologia Mediterranea0031-94651593-20952024-11-0163332333410.36253/phyto-1562314460Isolation, characterization and genomic analysis of a novel lytic bacteriophage infecting Agrobacterium tumefaciensMiloud SABRI0https://orcid.org/0000-0003-2574-4842Kaoutar EL HANDI1https://orcid.org/0000-0001-6293-2047Orges CARA2https://orcid.org/0009-0000-0682-0796Angelo DE STRADIS3https://orcid.org/0000-0003-1624-2365Toufic ELBEAINO4https://orcid.org/0000-0003-2211-7907International Centre for Advanced Mediterranean Agronomic Studies (CIHEAM of Bari), Via Ceglie 9, 70010 Valenzano (Ba), ItalyInternational Centre for Advanced Mediterranean Agronomic Studies (CIHEAM of Bari), Via Ceglie 9, 70010 Valenzano (Ba)International Centre for Advanced Mediterranean Agronomic Studies (CIHEAM of Bari), Via Ceglie 9, 70010 Valenzano (Ba)National Research Council of Italy (CNR), Institute for Sustainable Plant Protection (IPSP), University of Bari, Via Amendola 165/A, 70126 BariInternational Centre for Advanced Mediterranean Agronomic Studies (CIHEAM of Bari), Via Ceglie 9, 70010 Valenzano (Ba)Agrobacterium tumefaciens causes crown gall, and economic losses in important crops, including apple, pear, peach, and almond. Difficulties controlling this disease with conventional pesticides require alternative antibacterial agents. A novel lytic bacteriophage, Agrobacterium phage PAT1 (PAT1), with high lysis potential against A. tumefaciens, was isolated from wastewater. Interaction between PAT1 and A. tumefaciens cells was investigated using transmission electron microscopy. PAT1 adsorbed, infected, and replicated on A. tumefaciens in ≤30 min. Turbidity assays showed that PAT1 [Multiplicity of Infection (MOI) = 1] inhibited A. tumefaciens growth by 82% for 48 hours. PAT1 was resistant to broad ranges of pH (4 to 10) and temperatures (4 to 60°C). Bioinformatics analyses of the PAT1 genomic sequence showed that the bacteriophage was closely related to Atuphduovirus (Autographiviridae) phages. The PAT1 genome size was 45,040 base pairs with a G+C content of 54.5%, consisting of 54 coding sequences (CDS), of which the functions of 23 CDS were predicted, including an endolysin gene which could be used as an antimicrobial against A. tumefaciens. No lysogenic mediated genes or genes encoding virulence factors, antibiotic resistance, or toxins were detected in PAT1 genome. The bacteriophage showed potential as a biocontrol agent against A. tumefaciens infections, expanding the limited catalogue of lytic A. tumefaciens phages, although efficacy for control of crown gall in planta remains to be evaluated.https://oajournals.fupress.net/index.php/pm/article/view/15623plant pathogenic bacteriabiocontrolcrown gallphage therapy
spellingShingle Miloud SABRI
Kaoutar EL HANDI
Orges CARA
Angelo DE STRADIS
Toufic ELBEAINO
Isolation, characterization and genomic analysis of a novel lytic bacteriophage infecting Agrobacterium tumefaciens
Phytopathologia Mediterranea
plant pathogenic bacteria
biocontrol
crown gall
phage therapy
title Isolation, characterization and genomic analysis of a novel lytic bacteriophage infecting Agrobacterium tumefaciens
title_full Isolation, characterization and genomic analysis of a novel lytic bacteriophage infecting Agrobacterium tumefaciens
title_fullStr Isolation, characterization and genomic analysis of a novel lytic bacteriophage infecting Agrobacterium tumefaciens
title_full_unstemmed Isolation, characterization and genomic analysis of a novel lytic bacteriophage infecting Agrobacterium tumefaciens
title_short Isolation, characterization and genomic analysis of a novel lytic bacteriophage infecting Agrobacterium tumefaciens
title_sort isolation characterization and genomic analysis of a novel lytic bacteriophage infecting agrobacterium tumefaciens
topic plant pathogenic bacteria
biocontrol
crown gall
phage therapy
url https://oajournals.fupress.net/index.php/pm/article/view/15623
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AT orgescara isolationcharacterizationandgenomicanalysisofanovellyticbacteriophageinfectingagrobacteriumtumefaciens
AT angelodestradis isolationcharacterizationandgenomicanalysisofanovellyticbacteriophageinfectingagrobacteriumtumefaciens
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