A Tentative Stydy Of Application Of Volvariella Volvacea Opening Antisense Gene

The purpose of this study is to obtain a new Volvariella volvacea variety whose opening is inhebitated.In this study,with recipient strain Volvariella volvacea H104, we introduced pCAMBIA1301 plasmid,and got transformant H104 with hygromycin resistance and GUS activity.And,by calculating genetic transformation rate,we optimized agrobacterium C58 mediated transformation condition:cultivated mycelium for 3～4 days,infected when the concentration of agrobacterium is OD600 0.5～0.6 with 100μM AS in it,and then cocultivated for 5 days.With optimum condition,we introduced opn39,which carries anti-opning gene,and opn15,which carries opening gene,into H104,and got transformants VC58-opn39 and VC58-opn15.In fruiting experiment,inhabitation of opning of VC58-opn39 is observed,but we also observed that opning of VC58-opn15 is inhebitated unexpectly.That may result from homology between Volvariella volvacea genomic sequence and the sequence of the plasmid introduced,which subsequently caused nonspecific silencing.But we coundn’t bring to a conclusion without enough information from nucleic-aid database.For more information,we extracted genomic DNA and amplified the flanking sequence of opn gene(upstream 924 bp,downstream 500 bp,total 1106bp after merged)with TAILPCR and predicted by bioinformatics analysis.When analysed OFR,we found a ORF including opn gene fragment,which proved the correction of previous operation steps. After being analysed by blastn,we didn’t predict advanced structure of the 1106 bp fragment because of lacking of information from nucleic-aid database.Till now, exploration of Volvariella volvacea opening relative gene is not end yet,the follow-up work is still hard and hornerable.