secA gene suitability for fast and easy identification of Phytoplasmas by RFLP analysis

In this study we investigated the secA gene as possible marker to supplement the classification scheme based on 16S rRNA sequences. We cloned and sequenced secA gene and its flanking region of 'Ca. P. asteris' CPh strain phytoplasma, as well as 1997 bp length secA gene fragment of Canada peach X-disease CX and 1327 bp fragment of 'Ca. P. ziziphi' JWB. This helped us to select a primer pair for a single step PCR that can amplify similar to 1200 kb length fragment of secA gene from different phytoplasma groups. We propose that this fragment could be used in RFLP analysis to quickly identify and distinguish 16SrI-A, I-B, I-C, I-D, III-A, III-B, III-E, III-F, III-H, V-B, XII-B and XXI-A subgroup phytoplasmas using just two restriction endonucleases.

Automated summary

The study investigated the use of the secA gene as a marker to supplement the classification scheme based on 16S rRNA sequences. Cloning and sequencing of different phytoplasma groups' secA gene fragments allowed for the proposal of a method to quickly identify and distinguish phytoplasma subgroups.