Shotgun proteomic analysis of mulberry dwarf phytoplasma

Mulberry dwarf (MD), which is caused by phytoplasma, is one of the most serious infectious diseases of mulberry. Phytoplasmas have been associated with diseases in several hundred plant species. The inability to culture phytoplasmas in vitro has hindered their characterization at the molecular level. Though the complete genomes of two phytoplasmas have been published, little information has been obtained about the proteome of phytoplasma. Therefore, the proteomic information of phytoplasmas would be useful to elucidate the functional mechanisms of phytoplasma in many biological processes.

MD phytoplasmas, which belong to the 16SrI-B subgroup based on the 16S DNA analysis, were purified from infected tissues using a combination of differential centrifugation and density gradient centrifugation. The expressed proteome of phytoplasma was surveyed by one-dimensional SDS-PAGE and nanocapillary liquid chromatography-tandem mass spectrometry. A total of 209 phytoplasma proteins were unambiguously assigned, including the proteins with the functions of amino acid biosynthesis, cell envelope, cellular processes, energy metabolism, nucleosides and nucleotide metabolism, replication, transcription, translation, transport and binding as well as the proteins with other functions. In addition to these known function proteins, 63 proteins were annotated as hypothetical or conserved hypothetical proteins.

Taken together, a total of 209 phytoplasma proteins have been experimentally verified, representing the most extensive survey of any phytoplasma proteome to date. This study provided a valuable dataset of phytoplasma proteins, and a better understanding of the energy metabolism and virulence mechanisms of MD phytoplasma.