| The forming of fruiting body of lentinula edodes is the most complicated and the most important developmental process in the whole life history of lentinula edodes, must interact with environment, heredity, physiology and several other factors.Studying the developmentally related genes of fruiting body is always one of the most important questions in mushroom filed. Natural fruiting body of lentinula edodes differentiated into pileus, gill and stipe.Spherical Fruiting Body of lentinula edodes is a kind of mutated strain that was found in Sanligang town, Sui county, Hubei province, the whole fruiting body of this strain is spherical, just like a ball, the fruitbody is not opening, the stipe is not elongating and has no differentiation of gills.We isolated this strain and cultivated it again, the fruiting body is still spherical.This experiment used differential-display reverse transcription-PCR to investigate the difference of gene expression between mycelium, primordium and fruiting body of spherical fruiting body of lentinula edodes. The experiment used 3 anchor primers by 26 random primers to amplification. Only 14 random primers could get the stabled and clean bands. The cDNA which is transcribed by anchor primer PA and PC are better than anchor primer PG.Finally, we obtained 41 stabled fragments,17 fragments of them are positive fragments testified by reverse northern blotting, The false positive is 58.54%. Cloning and sequencing the 17 fragments, compared with Genbank datas by B,7 fragments had high homology with some known functional gene and protein sequence.Differential fragment GP20-540 had 98% homology with Lentinula edodes LephrA gene for photoreceptor A, it might encode a blue-light receptor, participating in biosynthesising of carotenoids in mycelia and resetting of circadian clock; differential fragment AP10-302 had 38% homology with Homing endonucleaseâ… -Apeâ…¡of uncultured crenarchaeote 45-H-12, it might be related to protein splicing; differential fragment CP16-324 had 43% homology with aspartokinase of Aspergillus terreus, it might encode aspartokinase, participating in biosynthesizing of lysine; differential fragment CP18-394 had 41% homology with Hsp90 co-chaperone Cdc37 of Aspergillus fumigatus,it might be related to stress tolerance and keeping the stability of proteins and activity of kinase as Chaperone; differential fragment GP7-502 had 50% homology with glycoside hydrolase family 2 protein of Laccaria bicolor,it might be related to glycometabolism;differential fragment AP21-479 had 60% homology with metallothionein of Ganoderma lucidum,it might be related to detoxification of heavy metals, transportation of metal ions and keeping them balance, ionic metabolization and cleaning active oxygen; differential fragment AP21-479 had 60% homology with hypothetical protein CC1G01848 of Coprinopsis cinerea, however,the function of this protein is unknown.Analysing the difference of gene expression between natural fruiting body and spherical fruiting body of lentinula edodes is the next phase of work,it will be much too meaningful for cloning and characterization of developmentally related genes from fruiting body of lentinula edodes. |
