TABLE 1.

Strains and plasmids used in this study

Strain or plasmidRelevant characteristicsaReference or source
X. campestris pv. vesicatoria
    85-10Pepper race 2; wild type; expresses avrBs1 and avrBs2; Rifr10
    85-10ΔhrpG85-10 derivative; carries a nonpolar hrpG deletion; Rifr45
    85-10ΔR85-10 derivative; carries a deletion from hpaF to hpaG; Rifr34
    85-10ΔL85-10 derivative; carries a deletion from ORF1 to xopA; Rifr34
    85-10ΔORF185-10 derivative; carries a deletion in ORF1; RifrThis study
    85-10ΔhpaH-oof85-10 derivative; carries a frameshift mutation in hpaH; RifrThis study
    85-10ΔxopA85-10 derivative; carries a deletion in xopA; RifrThis study
    85-10ΔxopD85-10 derivative; carries a deletion and a frameshift mutation in xopD; RifrThis study
    85*85-10 derivative containing the hrpG* mutation; constitutive hrp gene expression; Rifr45
    82*82-8 derivative containing the hrpG* mutation; constitutive hrp gene expression; pepper race 1; Rifr45
    85*ΔL85* derivative; carries a deletion from ORF1 to xopA; RifrThis study
    85*ΔhrpX85* derivative with a nonpolar hrpX deletion; Rifr34
    85*ΔhrcV and 82*ΔhrcV85* and 82* derivatives with a nonpolar hrcV deletion; nonfunctional TTSS; Rifr35, 36
    85*::pICxopA85* derivative containing pICxopA; expresses c-myc-tagged XopA; Rifr TcrThis study
    85*::pICxopD85* derivative containing pICxopD; expresses c-myc-tagged XopD; Rifr TcrThis study
    85-10::pICxopA85*::pICxopA derivative; wild-type hrpG restored using pOG; Rifr TcrThis study
    85-10::pICxopD85*::pICxopD derivative; wild-type hrpG restored using pOG; Rifr TcrThis study
E. coli
    DH10BF mcrA (mrr-hsdRMS-mcrBC) φ 80d lacZΔM15 ΔlacX74 endA1 recA1 deoR D(ara, leu)7697 araΔ139 galU galK λnupG rpsLBRL, Bethesda, Md.
    DH5αF φ80d lacZΔM15 Δ(lacZYA-argF)U169 endA1 recA1 deoR hdsR17 (rk mk+) phoA supE44 λ thi-1 gyrA96 relA1BRL
    DH5α λpirDH5α derivative, λpir; for replication of pIC1 and pOK derivatives32
Plasmids
    pUC118ColE1 replicon; Apr42
    pULB1pUC118 derivative with 7.5-kb BamHI insert from pXV331; contains ORF1 to xopD34
    pB-KS, PB-SKPhagemid, pUC derivative; AprStratagene
    pK2pB-KS derivative containing a 4.5-kb insert of pXV9; carries hrpA and 1.5-kb downstream region44
    pBLB1pB-KS derivative containing a 7.5-kb BamHI insert from pXV331; carries ORF1 to xopD34
    pBLH1pB-KS derivative containing a 3.0-kb HindIII insert from pXV331 (xopD)34
    pBFpB-KS derivative containing a 5.3-kb EcoRV insert from pXV2 containing hrpF23
    pBL5rpB-KS derivative containing a 2.3-kb EcoRV insert from pXV4 containing hpaF and hpaG23
    pRK2013Helper plasmid; ColE1 replicon TraRK+ Mob+; Kmr17
    pDS300FpDSK602 derivative carrying avrBs3 tagged with FLAG epitope at the C terminus; broad host range; Smr41
    pLAFR3RK2 replicon Mob+ Tra; contains plac; Tcr39
    pL3hpaFCpLAFR3 derivative carrying hpaF and hpaG; HpaF is c-myc tagged at amino acid position 66This study
    pXV331pLAFR3 derivative; contains a region from downstream of ORF1 to hrpA from X. campestris pv. vesicatoria strain 75-334
    pXV4pLAFR3 derivative; contains a region from hrpC2 to hpaG from X. campestris pv. vesicatoria strain 75-343
    pXV4::462Tn 3-gus insertion derivative of pXV4; Tcr Kmr43
    pXV9pLAFR3 derivative; contains region from ISX c7 to hrpE from X. campestris pv. vesicatoria strain 75-39
    pOKSuicide vector; pKNG101 derivative; Smr/Sucs24
    pOGpOK derivative; carries wild-type hrpG34
    pOLBpOK derivative; carries a 3.7-kb insert with a 3.8-kb deletion from ORF1 to xopA34
    pORpOK derivative; carries a 5.7-kb insert with a 2.5-kb deletion from hpaF to hpaG34
    pOORF1pOK derivative; carries a 4.4-kb insert with a 1.0-kb deletion in ORF1This study
    pOhpaHpOK derivative; carries a 5.6-kb insert with an out-of-frame mutation at codon position 38 of hpaHThis study
    pOxopApOK derivative; carries a 2.4-kb insert with a 0.3-kb deletion in xopAThis study
    pOxopDpOK derivative; carries a 1.85-kb insert with a 0.15-kb deletion in xopD causing an out-of-frame mutation after codon position 311 of xopDThis study
    pIC1Integrative vector; carries a triple-c-myc epitope followed by a promoterless uidA gene; Tcr34
    pICxopApIC1 derivative; carries the entire xopA ORF fused in frame to the c-myc epitopeThis study
    pICxopDpIC1 derivative; carries the entire xopD ORF fused in frame to the c-myc epitopeThis study
  • a Ap, ampicillin; Km, kanamycin; Rif, rifampin; Sm, spectinomycin; Suc, sucrose; Tc, tetracycline; r, resistant; s, sensitive.