1 Cloning vector lacZ, Kmr, Ampr, Invitrogen, California, USA    pBBR1MCS-5 Cloning vector, Gmr [29]    https://www.selleckchem.com/products/PLX-4032.html pMP220 Promoter-probe vector containing a promoterless lacZ gene [30]    pCR::ORF0 integrative plasmid pCR2.1 carrying ORF0 This

study    pCR::ORF1 integrative plasmid pCR2.1 carrying ORF1 This study    pCR::ORF2 integrative plasmid pCR2.1 carrying ORF2 This study    pCR::mgoB integrative plasmid pCR2.1 carrying mgoB This study    pCR::mgoC integrative plasmid pCR2.1 carrying mgoC This study    pCR::mgoA integrative plasmid pCR2.1 carrying mgoA This study    pCR::mgoD integrative this website plasmid pCR2.1 carrying mgoD This study    pCG2-6 genomic clone of UMAF0158 GenBank-DQ532441 [15]    pLac36 From mgoB to mgoD

cloned in pBBR1MCS-5 This study    pLac56 mgoA and mgoD cloned in pBBR1MCS-5 This study    pLac6 mgoD cloned in pBBR1MCS-5 This study    pMPmgo pMP220 vector containing the putative promoters of mgo operon This study    pEMG integrative plasmid for deletion mutagenesis, Kmr. [31]    pSW-2 plasmid carrying I-SceI gene for deletion mutagenesis, Gmr. [31] a) Ampr: ampicillin resistance; Gmr: gentamycin resistance; Kmr: kanamycin resistance; Nfr: Nitrofurantoin resistance. b) CECT: Spanish Type Culture Collection. c) ORF0 was named in this way because it was cloned as an uncompleted ORF LXH254 manufacturer Detection of P. syringae toxin production Syringomycin complex production by strains of P. syringae strains was detected using growth inhibition tests performed on potato dextrose agar (PDA) against Geotrichum candidum [32] and nutrient agar against Rhodotorula pilimanae [33]. Mangotoxin production was assayed using the indicator technique, which has been described previously and involves growth inhibition of Escherichia coli on

Pseudomonas next Minimal Medium (PMS [34]). Briefly, a double layer of the indicator microorganism was made with E. coli CECT831. After solidification, the P. syringae wild-type strain and its derivatives mutants were stabbed, and the plates were incubated at 22°C for 24 h, followed by an additional 24 h at 37°C. To determine the identity of the biochemical step that is putatively targeted by mangotoxin, the same plate bioassay was performed in separate plates with the addition of 100 μl of a 6 mM solution of ornithine or N-acetyl-ornithine. To assess the production of mangotoxin in liquid cultures, we used a cell-free filtrate dilution as previously described [13]. Insertional inactivation mutagenesis Insertional inactivation mutagenesis of P. syringae pv.