The Studies On Identification,Metabolites Separation Andbioactivity Of Endophytic Fungi DYSJ3 From Aphanamixis Grandifolia

Aphanamixis grandifolia is a traditional Chinese medicine distributed across the tropical and subtropical areas.Now the study on A.grandifolia was mainly on the extraction and application of active products,and there was few study on its endophyte and bioactive metabolites.An endophytic fungi strain DYSJ3 was screened from Aphanamixis grandifolia,which had obvious inhibition effects to 4 of plant pathogenic fungi(Fusarium oxysporum f.sp.Cubense?Colletotrichum musae?Colletotrichum gloeosporioides(Penz.)Sacc?Colletotrichum gloeosporioides Penz).All the inhibitory rate could be up to 50%.Strain DYSJ3 was identified as Aspergillus versicolor by morphological and molecular biological methods.Strain DYSJ3 was fermented in liquid culture medium,and mycelium was disrupted by ultrasonic crushing method,extracted by ethyl acetate.The crude extractings were stability to heat and visible light.After 1 hour of temperature 121 ?,the inhibition effect had no significant changes.While exposed to visible light for 14d,the extract showed no decrease in activity.When the concentration of the extract was 1000?g/mL,the inhibitory ratio to Colletotrichum musae?Botryosphaeria berengriana f.sp.Piricola?Fusarium oxysporum f.sp.Cucumerinum?Colletotrichum gloeosporioides(Penz.)Sacc?Gibberella zeae was all exceeded 90%.The extract especially has better inhibition effects to Colletotrichum gloeosporioides(Penz.)Sacc and Botryosphaeria berengriana f.sp.Piricola,with the inhibitory ratio was 93.5%and 87.3%when the concentration is 500?g/mL.The compounds from the extract were isolated by column chromatography combined with bioactivity-gudied method.Eight compounds were isolated from mycelium extract(A-H).The structures of all the compounds were elucidated by spectral data.They were elucidated as:A:Averufanin;B:Ergosterol peroxide;D:Versicolorin B;G:Averythrin;H:Sterigmatocystin.The activites of eight compounds to Colletotrichum musae and Colletotrichum gloeosporioides were evaluated by spore germination and paper filtering method.Compound D and H showed inhibitory activity to Colletotrichum musae,and compound D also exhibited inhibition effects to spore germination of two pathogens with the EC50 was 0.llmg/mL and 0.07mg/mL,respectviely.D also showed effects on controlling banana anthracnose and mango anthracnose.