Inhibitory Effect Of Berberine On Two Important Plant Pathogenic Fungi And Preparation Of Berberinr Fumigant

Apple ring rot and peach brown rot are severe diseases caused by Botryosphaeria dothidea and Monilinia fructicola,respectively.The two diseases give rise to considerable economic loss in agriculture.As a botanical fungicide,berberine is pretty promising in future applications.In present study,experiments involve three aspects:efficacy evaluation of berberine and its compounds,exploration of berberine targrts,and preparation of berberine fumigant to investigate the inhibitory effect of berberine on B.dothidea and M.fructicola,as well as development of new dosage form of berberine.1.The EC50 values of berberine,405-1,405-2,procymidone and difenoconazole against B.dothidea were 8.85 ?g/mL,5.45 ?g/mL,2.42?g/mL,0.55 ?g/mL and 0.086 ?g/mL,respectively.And the EC50 values of these five fungicides against M fructicola were 8 ?g/mL,1.35 ?g/mL,0.59?g/mL,0.35 ?g/mL and 0.0635?g/mL,respectively.For B.dothidea,synergistic effect occurred when the ratio of berberine to 405-1 was 5:1.For M.fructicola,it occurred when the ratio of berberine to 405-2 was 5:1.2.It was found that ENO was candidate target of berberine against M.fructicola.Hence,the ENO gene was cloned.The coding sequence(CDS)of the gene was 1317 bp,which was an intact ORF encoding 438 amino acids.In addition,recombinant Escherichia coli pET-28a-EVO/BL21 was correctly constructed,and the size of its expression band was consistent with the predicted molecular weight.3.Recombinant Pichia pastoris pPIC9K-ENO/GS 115 and filamentous fungal expression vector were successfully constructed,which laid a foundation for verifying whether ENO was a target of berberine against M.fructicola.4.Berberine fumigant was well prepared,and it was composed of 10%berberine,30%fuel A,45%oxidant B,10%smoke agent C,and 5%temperature-lowering agent D.Its physicochemical indexes were determined in view of the fumigant product standard HG-T2467.19-2003.