Proteome analysis of rice blast fungus (Magnaporthe grisea) proteome during appressorium formation.
We used two-dimensional gel electrophoresis (2-DE) to identify the proteins that are induced in the rice blast fungus Magnaporthe grisea during appressorium formation. Proteins were extracted from conidia that had germinated on hydrophilic glass plates or from germinated and appressoria-forming conidia on leaf wax-coated hydrophobic glass plates after 4, 8, and 12 h of incubation. Differentially expressed protein spots during appressorium formation were confirmed from gels after 2-DE analysis where proteins had been labeled with (35)S methionine and stained with silver. Internal amino acid sequencing identified five proteins among several proteins induced during appressorium formation. Two denoted as M. grisea proteasome homolgues (MgP1 and MgP5) were 20S proteasome alpha subunits. The remaining three were scytalone dehydratase (SCD), and serine carboxypeptidase Y (CPY). None of the five have been reported previously in the rice blast fungus apart from SCD. We further investigated the role the alpha subunit of 20S proteasome plays in appressorium formation. We confirmed by Western blot analysis that MgP5 is highly expressed during appressorium formation and found that it is also markedly induced by nitrogen- and carbon-starvation, in particular by the former. These observations suggest that the 20S proteasome may be involved in remobilizing storage proteins, which then help to build the appressorium. Thus, fungal proteome analysis may provide important clues about developmental changes such as the generation of the appressorium.
Kim
Sun Tae
ST
Division of Applied Life Science, Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Chinju, Korea.
Yu
Seok
S
Kim
Sang Gon
SG
Kim
Han Ju
HJ
Kang
Sun Young
SY
Hwang
Du Hyeon
DH
Jang
Yu Sin
YS
Kang
Kyu Young
KY
D000595:Amino Acid Sequence; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D005656:Fungal Proteins / Q000378:metabolism*; D020869:Gene Expression Profiling; D020082:Magnaporthe / Q000254:growth & development / Q000378:metabolism*; D008969:Molecular Sequence Data; D010802:Phylogeny; D020543:Proteome / Q000378:metabolism*; D013997:Time Factors
D005656:Fungal Proteins; D020543:Proteome