| Pleurotus ostreatus, a model organism for studying Pleurotus, is one of the four major cultivated mushroom species. It attracts people not only for its delicious flavor, but also for its medicinal ant economic values. A complete life cycle of Pleurotus ostreatus includes five separate stages:basidiospore, basidiospore, primordium and fruiting body. Of all the stages, its morphology changes most in the development of dicaryon mycelia knotting into the primordium form. However, the mechanism of this change remains unclear, presumable cause is that it regulates metabolism by ion concentration, enzyme activity and gene expression. This study focused on the differentially expressed genes between mycelium and primordium in Pleurotus ostreatus, which is significant to the study of its differentiation and breeding.Mycelium M739 and primordium P739 were used as the material in this study, which both come from the strain Po739 kept in our lab. In order to find out differentially expressed genes between mycelium and primordium, suppression subtractive hybridization was used to construct both forward and reverse subtractive library. Colony PCR showed that cDNA segments tested in forward and reverse subtractive library were among 250 bp-1000 bp and that the effective recombination rates reached 85.4% and 83% respectively. By reverse Northern blotting,200 obviously color-gap clones were selected from both libraries to sequence.By homology comparison in Genbank database and function classification in Gene ontology,69.5% and 66.5% of the genes were annotated, which involve in various biochemical processes such as catalytic activity, enzyme activity, signal transduction, regulation of translation, cellular pathways, biological channels. Then genes encoding mitochondrial-processing peptidase and voltage-dependent anion channel were selected to apply RT-PCR to testify their differentially expression and electrophoresis confirmed the results in reverse Northern blotting. |
PDF Full Text Request