Shotgun proteomic analysis of mulberry dwarf phytoplasma
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Published on: August 27, 2010
Xianling Ji1, Yingping Gai2, Baoyun Lu1
1College of Forestry, Shandong Agricultural University, Tai'an, Shandong, 271018, PR China.
Background:
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.
Results:
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.
Conclusions:
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.
Insights
This study identified 209 mulberry dwarf (MD) phytoplasma proteins, offering insights into phytoplasma energy metabolism and virulence. This proteomic survey advances understanding of this significant plant pathogen.
Area of Science:
- Plant Pathology
- Molecular Biology
- Proteomics
Background:
- Mulberry dwarf (MD) is a serious disease caused by phytoplasma, affecting mulberry and other plants.
- Difficulty in in vitro culturing hinders phytoplasma molecular characterization.
- Limited proteomic data exists despite sequenced genomes, impeding functional mechanism understanding.
Purpose of the Study:
- To characterize the proteome of mulberry dwarf (MD) phytoplasma.
- To identify proteins involved in MD phytoplasma's biological processes.
- To gain insights into MD phytoplasma's energy metabolism and virulence.
Main Methods:
- Purification of MD phytoplasmas from infected mulberry tissues using centrifugation techniques.
- Proteomic analysis via one-dimensional SDS-PAGE and nanocapillary LC-MS/MS.
- Bioinformatic assignment of identified proteins based on known functions and hypothetical status.
Main Results:
- Successfully identified and assigned 209 unique phytoplasma proteins.
- Detected proteins involved in essential cellular functions like metabolism, replication, and transport.
- Annotated 63 hypothetical or conserved hypothetical proteins, expanding the known protein repertoire.
Conclusions:
- This study presents the most extensive phytoplasma proteome survey to date.
- Provides a valuable protein dataset for MD phytoplasma research.
- Enhances understanding of MD phytoplasma's energy metabolism and virulence factors.
