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The protein samples were separated by two-dimensional gel electrophoresis and stained
with Coomassie Brilliant Blue 250. Proteins were isolated using to a phenol-based
extraction procedure (Hurkman and Tanaka 1986). Ploidy level was determined by flow
cytometry using two step procedure with Otto I+II buffers and the fluorochrome DAPI.
Results
Although in previous studies using PAGE technique we have not
observed significant differences between gametophyte and sporophyte,
the patterns obtained by two-dimensional gel electrophoresis showed
the existence of distinct spots for both gametophyte and sporophyte
extracts. The differences in protein expression suggest that gametophyte
and sporophyte have different metabolic pathways of protein synthesis.
G1 G2 G3 G4 G5 S1 S2 S3 S4 S5
gametophyte sporophyte
Athyrium filix-femina
Results
gametophyte sporophyte
Asplenium trichomanes
Results
gametophyte sporophyte
Polypodium vulgare
Conclusions
This study presents a preliminary analysis of proteomics profiles and compares the ploidy
level for detection of further changes that can occur at the molecular level during the
cryopreservation process. Proteomic analysis allows the observation of translationally
manifested cell physiological response.