These two genomic regions are shown in Fig. 1 as white boxes. Both genomic fragments were successfully amplified from all 26 cultivars, and sequencing analysis confirmed that all 26 strains harbored dnaD, imp, and idpA. These two region sequences have been deposited in the GenBank database: AB636356-407.

Analysis of the deduced amino acid sequences using sosui ver 1.11 (Hirokawa et al., 1998) yielded the predictions that Imp contains a transmembrane region in its N-terminal region and a hydrophilic domain, and that IdpA contains both N- and C-terminal transmembrane regions, as well as a central hydrophilic domain (Fig. 1b). These features are identical to those of other previously analyzed Imp and IdpA proteins (Kakizawa INK 128 mw et al., 2006a, 2009). Analysis of the Imp and IdpA sequences using the SignalP program with the hidden Markov model yielded the relatively high-probability predictions (0.885 and 0.824, respectively)

that the two proteins have signal sequences. The signal sequence cleavage sites were predicted to lie between amino acid residues 48 and 49 for Imp, and between residues 35 and 36 for IdpA. Analysis of the Imp and IdpA sequences using the Psort program suggested with low probability (0.300) that Imp may be secreted from the bacterial cell and with high probability that IdpA is an integral membrane protein. BEZ235 clinical trial There were no silent substitutions in the PoiBI imp genes. Of the 26 PoiBI Imp amino acid sequences obtained from the 26 PoiBI-infected cultivars, those from ‘Annette Hegg Maxi’, ‘Annette Hegg Pink’, ‘Annette Hegg Supreme’, ‘Arctic’, ‘Jingle Bells’, ‘Premium Red’, and ‘Winter Rose White’ were 100% identical, and those from ‘Prestige Bright Red’, ‘Primero Jingle Bells’, and ‘Vision of Grandeur’ were identical. Therefore, in comparing the encoded Imp amino acid sequences, we used those from ‘Winter Rose White’ and ‘Primero Jingle Bells’, respectively, to represent these two groups

of identical sequences. The resulting multiple alignment of these sequences and that of WX Imp is shown in Fig. 2a. Although variations in the PoiBI Imp sequences were noted at several positions, the sequence identity was overall very high. The lowest sequence identity score (97.2%) was obtained for the comparison of ‘Enduring Pink’ vs. ‘Jester Jingle Bells’, ‘Jester Marble’, and ‘Peterstar Vitamin B12 Marble’. A phylogenetic tree of the PoiBI Imp amino acid sequences is shown in Fig. 2b. In contrast to the diversity of imp genes, there was no difference in the sequences of the 16S rRNA gene, idpA, or dnaD genes from the 26 poinsettia cultivars. The amino acid sequences deduced from PoiBI and WX imp, idpA, and dnaD are shown in Fig. S1. Among the PoiBI and WX Imp amino acid sequences, identity scores ranged from 92.6% to 93.8%, with a mean identity of 93.3%. The PoiBI and WX amino acid sequences of DnaD and IdpA had identity scores of 98.0% and 64.