The Effect Of Hypocrellin A On Cell Membrane And Antioxidant Enzymes Of Botrytis Cinerea

Hypocrellin A（HA）is a nature photosensitizer from Shiraia bambuscola and Hypocrella bambusea,at present it has been widely applied in medical field.Because of it has high insecticidal and bacteriostatic activity,and was safety to the environment,HA has a high developing potential to be developed as a photoactivated pesticide.The effect of Hypocrellin A on cell membrane and antioxidant enzymes of Botrytis cinerea was investigated.The main results were as following:1.Using Confocal laser scanning microscope（CLSM）,found that HA can be absorbed by the hyphae of Botrytis cinerea.2.Using TBA method to determine the content of MDA,found that HA can induce Botrytis cinerea lipid peroxidation.When the hyphae was treated with 40 mg.L^-1 HA,MDA content was increased with the treatment time prolonged.When the hyphae was treated with different concentration of HA,MDA content increased with the concentration of HA raised.The longer treatment time was and the higher concentration of HA,the more obvious the lipid peroxidation of hyphaee was.3.Using biuret reaction,anthrone colorimetry method,and uv spectrophotometry to determine the content of protein,soluble sugar,and nucleic acid,found that HA can change cell membrane permeability of Botrytis cinerea,resulting in leakage of the inclusions.When the hyphae was treated with 40 mg.L^-1 HA,the content of protein and soluble sugar of hyphae were decreased with the treatment time prolonged.When the hyphae was treated for 36 h,comparing with control group,the content were reduced by 37.0% and 55.2% respectively,while the content of extracellular nucleic acid and protein were increased.When the hyphae was treated with different concentration of HA,the content of protein and soluble sugar of hyphae were decreased with the concentration of HA raised.Comparing with control group,the content were reduced by 81.0% and 77.2% respectively,while the content of extracellular nucleic acid and protein were increased when the concentration of HA was 100 mg.L^-1.The longer treatment time was and the higher concentration of HA,the more obvious the cell membrane permeability changed.4.Using ATPase kit to determine ATPase activity of Botrytis cinerea,found that HA can inhibit ATPase activity.When the hyphae was treated with 40 mg.L^-1 HA,Na⁺,K⁺-ATPase and Ca²⁺,Mg²⁺-ATPase activity was increased at first and then decreased with the treatment time prolonged.When the hyphae was treated for 36 h,comparing with control group,ATPase activity were reduced by47.0% and 53.4% respectively.When the hyphae was treated with different concentration of HA,Na⁺,K⁺-ATPase and Ca²⁺,Mg²⁺-ATPase activity was increased at first and then decreased with the concentration of HA raised.Comparing with control group,ATPase activity were reduced by 66.1% and69.1% respectively,when the concentration of HA was 100 mg.L^-1.The longer treatment time was and the higher concentration of HA,the more obvious the enzyme activity inhibited.5.Using xanthine oxidase method,uv spectrophotometry,and guaiacol colorimetric method to determine SOD、CAT、POD activity of Botrytis cinerea,found that HA can inhibit antioxidant enzymes activity of Botrytis cinerea.When the hyphae was treated with 40 mg.L^-1 HA,SOD、CAT、POD activity was decreased with the treatment time prolonged.When the hyphae was treated for 36 h,comparing with control group,the activity were reduced by 27.7%、86.9% and 53.4% respectively.When the hyphae was treated with different concentration of HA,SOD 、CAT、POD activity was decreased with the concentration of HA raised.Comparing with control group,the activity were reduced by 30.9%、91.2%and 83.0% respectively,when the concentration of HA was 100 mg.L^-1.The longer treatment time was and the higher concentration of HA,the more obvious the antioxidant enzyme activity inhibited.The research found that HA can injure the membrane permeability,and inhibit antioxidant enzymes activity of Botrytis cinerea.It provide theory gist to develop HA as a photoactivated pesticide.