Identification of genetic markers to distinguish the virulent and avirulent subspecies of Pantoea stewartii by comparative proteomics and genetic analysis.
Pantoea stewartii subsp. stewartii (Pnss), the causal agent of Stewart's bacterial wilt and leaf blight of maize and sweet corn, is one of the quarantine pathogens in many countries and regions. In contrast, P. stewartii subsp. indologenes (Pnsi), the closely related subspecies of Pnss, is avirulent on these plants. In this study, the protein expression profiles of these two subspecies were compared using two-dimensional gel electrophoresis analysis. Twenty-one unique protein spots consistently detected in Pnss but not in Pnsi were analyzed by mass spectrometry. Some of these Pnss-specific proteins are known to be essential for virulence and survival in host, such as FoxR and HrcJ, which are the key components of iron uptake and Type III secretion systems, respectively. For further genetic analysis, six Pnss-specific proteins were characterized by peptide sequencing. Southern and Northern blot analyses revealed that the differences in protein expression profiles of the two subspecies were either due to the discrepancy at genome level or because of the variations in transcriptional expression. The results provide novel genetic markers to distinguish the two closely related subspecies and may also serve as useful clues for investigation of the genetic basis accounting for their sharp difference in virulence.
Wu
Qiong
Q
Institute of Molecular and Cell Biology, 61 Biopolis Drive, Singapore 138673, Singapore.
Jiang
Zide
Z
Liao
Jinliang
J
Chen
Zhinan
Z
Li
Huaping
H
Mei
Mantong
M
Zhang
Lian-Hui
LH
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015152:Blotting, Northern; D015139:Blotting, Southern; D015180:Electrophoresis, Gel, Two-Dimensional; D015964:Gene Expression Regulation, Bacterial; D005819:Genetic Markers; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D020636:Pantoea / Q000145:classification / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D010935:Plant Diseases / Q000382:microbiology; D018515:Plant Leaves / Q000382:microbiology; D040901:Proteomics; D020539:Sequence Analysis, Protein; D013045:Species Specificity; D014774:Virulence; D003313:Zea mays / Q000382:microbiology*
D001426:Bacterial Proteins; D005819:Genetic Markers