Analysis of Gene Expression Differences in the Substrate-Decomposing Ability Degenerated Strains of Agaricus bisporus

摘要

To investigate the gene expression differences and molecular mechanism of substrate-decomposing ability degeneration in Agaricus bisporus (J. E. Lange) Imbach, the strain As2796, cultivated most widely in China, as well as two substrate-decomposing ability degenerated mutants derived from which, 2796-3 and 2796-5, were analyzed by using the techniques of mRNA differential display reverse transcription-PCR ( DDRT-PCR) and two dimensional electrophoresis (2-DE). As a result, five special genes showing obviously differential expressions were found in DDRT-PCR, and some of them have high homologies with substratedecomposing enzymes. Based on the genome sequence data of A. bisporus, PCR primers were designed, and the coding sequences (CDS) with introns and about 1200 bp upstream regulative sequences of above five special genes were amplified, cloned and sequenced, but the comparison of the sequences among three strains showed that the genes and upstream sequences have no repeatable difference. In the 2-DE analysis, two proteins were found to be up-and down-regulated expression, respectively, with obvious and repeatable profiles in both degenerated strains. Based on the results of MALDI-TOF/MS analysis and database searching, these two proteins were identified as an actin and a putative NADH dehydrogenase iron-sulfur protein 3, respectively.