TABLE 1.
Strains, plasmids, and primer used in this study
| Strain, plasmid, or primer | Relevant characteristics | Reference or source |
|---|---|---|
| Escherichia coli strains | ||
| DH5α | supE44 ΔlacU169 (φ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | 43 |
| S17-1 λ-pir | λl-pir lysogen of S17-1 (thi pro hsdR hsdM+recA RP4:2− Tc::MU-Km::Tn7) Tcr Smr | 12 |
| Pseudomonas syringae strains | ||
| P. syringae pv. glycinea 1a/96 | Wild type isolated from soybean; Pvd+ Ach+ | 47 |
| P. syringae pv. syringae 22d/93 | Wild type isolated from soybean; Pvd+ Ach+ | 48 |
| P. syringae pv. syringae 22d/93ΔPvd | pvsA mutant carrying Kmr cassette in pvsA gene; Pvd− Ach+ Kmr | This study |
| P. syringae pv. syringae 22d/93ΔAch | acsD mutant carrying Kmr cassette in acsD gene; Pvd+ Ach− Kmr | This study |
| P. syringae pv. syringae 22d/93ΔSid | Tn5 mutant of P. syringae pv. syringae 22d/93ΔPvd carrying mini-Tn5 in acsD gene; Pvd− Ach− Kmr Spr | This study |
| Dickeya dadantii strains | ||
| 3937 cbsE-1 acsA-37 | cbsE::Ω acsA-37::MudII1734; chrysobactin− Ach− Spr Smr Kmr | 14 |
| 3937 acr-1 fct-34 | acr-1::Ω fct-34::lacZ; chrysobactin− Ach− Spr Smr Kmr | 16 |
| 3937 cbsE-1 tonB60 | cbsE::Ω tonB60::MudII1734; chrysobactin− Ach+ Spr Smr | 14 |
| Plasmids | ||
| pGEM-T Easy | TA cloning vector; Apr | Promega |
| pPVSA1 | 1.4-kb PCR fragment located in pvsA gene of P. syringae pv. syringae 22d/93, cloned in pGEM-T Easy; Apr | This study |
| pPVSA2 | 1.6-kb PCR fragment located in pvsA gene of P. syringae pv. syringae 22d/93, cloned in pGEM-T Easy; Apr | This study |
| pPVSA3 | 1.4-kb SpeI-KpnI fragment from pPVSA1 cloned in SpeI-KpnI-digested pPVSA2; Apr | This study |
| pPVSA4 | 1.8-kb KpnI Kmr cassette from pMKm cloned in KpnI-digested pPVSA3; Apr Kmr | This study |
| pACS1 | 1.3-kb PCR fragment located in acsF gene of P. syringae pv. syringae 22d/93, cloned in pGEM-T Easy; Apr | This study |
| pACS2 | 2-kb PCR fragment located in acsE and yhcA genes of P. syringae pv. syringae 22d/93, cloned in pGEM-T Easy; Apr | This study |
| pACS3 | 1.8-kb PstI Kmr cassette from pMKm blunt ended and cloned in SpeI-digested and blunt-ended pACS1; Apr Kmr | This study |
| pACS4 | 3-kb ScaI-EcoRI fragment from pACS3 blunt ended and cloned in EcoRV-digested pACS2; Apr Kmr | This study |
| pBBR1MCS | Broad-host-range cloning vector; Cmr | 30 |
| pBBR-11-Sid | 6-kb PstI fragment of P. syringae pv. syringae 22d/93ΔSid carrying Tn5 insertion site in pBBR1MCS; Cmr Spr | This study |
| pMKm | Donor of kanamycin cassette; Kmr | 41 |
| pUT/mini-Tn5 Sm/Sp | Mini-Tn5 encoding Smr/Spr on broad-host-range suicide plasmid pUT; Apr Smr Spr | 12 |
| Primers | ||
| pvsA_fwd1_Spe | 5′-AATACTAGTGGATCCTGATGCGACTGGCCTTCGATC-3′ | This study |
| pvsA_rev6_Kpn | 5′-TAAGGTACCACGTCGAGGCTGAGCGGATC-3′ | This study |
| pvsA_fwd7_Kpn | 5′-TTAGGTACCTCGAACTTGGCCTCGCGGCTG-3′ | This study |
| pvsA_rev4_Bam | 5′-TTTGGATCCGGCAGACCGTGGCTGAG-3′ | This study |
| Achr1_fwd | 5′-AGCGAGGACTCACAGATGTTG-3′ | This study |
| Achr2_KpnI_rev | 5′-GGTACCCAATGCTGCTGAATGGCAAC-3′ | This study |
| Achr5_EcoRV_fwd | 5′-GATATCAACTATGTGCGTCTTGCGTC-3′ | This study |
| Achr6_rev | 5′-ACGAATGCCACCAGACAGG-3′ | This study |