Table 2 Bacterial strains and plasmids used in this study

NEB10β

Δ(mrr-hsdRMS-mcrB) deoR recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ⁻ rpsL nupG

New England Biolabs

S17–1

Strain used to transfer pDR1 plasmid

[43]

NCPPB 3335

Pathotype strain, isolated from a diseased olive tree; contains three native plasmids (pPsv48A, pPsv48B and pPsv48C)

[78]

Psv48ΔAB

NCPPB 3335 derivative, cured of pPsv48A and pPsv48B

[18]

UPN25.1

UPN827 derivative containing pPsv48CΔ25, a 5.5 kb spontaneous deletion derivative of pPsv48C that spans the RepJ replicon, with no TA systems.

[37]

UPN508

Derivative of NCPPB 3335 containing pPsv48A::Tn5-GDYN1

[18]

UPN827

Derivative of Psv48ΔAB containing plasmid pPsv48C::Tn5-GDYN1

[18]

UPN864

UPN827 derivative, which contains plasmid pPsv48CΔ1

This work

UPN912

Plasmidless derivative of strain NCPPB3335; was obtained by curing pPsv48C::sacB from strain UPN1007

[35]

UPN1007

Derivative of Psv48ΔAB containing plasmid pPsv48C::sacB

This work

B728a

Plasmidless bean pathogen Cu^R, Rif^R, Sm^R

[79]

Plasmids

pBlueScript II SK

E. coli cloning vector; 2.96 kb, Amp^R

Stratagene

pBAD24

E. coli expression vector, contains a P_BAD promoter inducible with arabinose; 4.5 kb, Amp^R

[76]

pDR1

Delivery vector for Tn5-GDYN1, based on pSUP2021; Km^R, Gm^R, Sm^R, Sp^R; confers sucrose-dependent lethality

[43]

pGEM-T Easy

E. coli cloning vector; 3 kb, Amp^R

Promega

pJET1.2

E. coli cloning vector 2.9 kb, Amp^R

Thermo Fisher Scientific

pK18mobsacB

Mobilizable cloning vector, confers sucrose-dependent lethality; Km^R, Suc^S

[71]

pKMAG

E. coli vector derived from pK184, devoid of the Plac promoter and containing the transcriptional terminator and polylinker from pME6041; 2.6 kb, Km^R; accession no. KX714576

[37]

pKMAG-C

pKMAG containing the minimal RepA-PFP replicon from pPsv48C, replicates in E. coli and in Pseudomonas; 4.3 kb, Km^R; accession no. KX714577

[37]

pME6041

Broad host range cloning vector; 5.6 kb, Km^R

[80]

pPsv48A

Virulence plasmid of NCPPB 3335 (accession n° FR820585); with RepA-PFP replicon; 80.1 kb

[18]

pPsv48A::Tn5-GDYN1

Plasmid pPsv48A tagged with Tn5-GDYN1 at position 1469

[18]

pPsv48B

Native plasmid of NCPPB 3335 (accession no. FR820586); with RepA-PFP replicon; 45.2 kb

[18]

pPsv48C

Virulence plasmid of NCPPB 3335 (accession no. FR820587); with RepA-PFP and RepJ replicons; 42.1 kb

[18]

pPsv48C::sacB

pPsv48C derivative containing the Km^R-sacB cassette from pK18mobsacB inserted at position 26,916; Km^R, Suc^S

This work

pPsv48C::Tn5-GDYN1

pPsv48C containing Tn5-GDYN1 inserted at position 37,036; Km^R, Gm^R, Suc^S

This work

pPsv48CΔ1

Derives from pPsv48C::Tn5GDYN1 by the spontaneous deletion of 8.3 kb (positions 32,807–41,121 of FR820587) mediated by recombination between two copies of MITEPsy2

This work

pPsv48CΔ25

Spontaneous sucrose-resistant deletion derivative from pPsv48C::sacB, generated by a one-ended transposition of IS801; this plasmid is 5.5 kb long, spanning positions 27,019–32,557 of FR820587

[37]

pRK3A

pRK415 derivative containing genes PSPSV_A0043, PSPSV_A0032 and PSPSV_A0020 cloned in tandem in this order, each with their own promoter and under the control of the P_lac promoter from the vector; Tc^R

This work

pRK3C

pRK415 derivative containing genes PSPSV_C0050, PSPSV_C0008 and PSPSV_C0003 cloned in tandem in this order, each with their own promoter and under the control of the P_lac promoter from the vector; Tc^R

This work

pRK415

Broad host range cloning vector; 10.5 kb, Tc^R

[81]