Study On The Anti-Phytopathogen And Antioxidant Activities Of Secondary Metabolites From Gymnospermium Microrrhynchum (S. Moore) Takht

Natural products play an irreplaceable role in food,medicine,agriculture and reletive fields.Berberidaceae plants,famous for their rich in active substrate,making it reasonable for us to choose G.microrrhynchum included in Berberidaceae plants to get more from it.This research reported the extraction and isolation of secondary metabolites from G.microrrhynchum,and the exploration of the anti-phytopathogenic and antioxidant activity of these compounds.The roots and stems of G.microrrhynchum were ground into powder,and the crude extracts were obtained by ultrasonic-assisted leaching.The Inhibitory activity of root and stem extract from G.microrrhynchum to Setosphaeria turcica,Magnaporthe oryza,Fusarium graminearum,Botrytis cinerea was assisted by growth rate method.Ningnanmycin was selected as positive control,anti-Tobacco Mosaic Virus activity of crude root extract of G.microrrhynchum was evaluated by half leaf spot method.The results showed that the root and stem extracts of G.microrrhynchum exhibited inhibitory effect against Magnaporthe oryza with the inhibition rates were 46%and 37%,respectively.The crude extract of peony grass root can reduce the number of dead spots caused by tobacco mosaic virus,and the inhibition rate of the incidence rate is 21.21%.Antioxidant activity of peony root and stem extracts of G.microrrhynchum were evaluated by DPPH free radical scavenging method.As it tells,they comes to good antioxidant activity,and the root extract is more infectous.Three-part polar extract(petroleum ether phase,ethyl acetate phase,n-butanol phase)are got from root extract of G.microrrhynchum by organic solvent fractional extraction.Their antioxidant activity was evaluated by the DPPH free radical clearance method,the antioxidant activity of ethyl acetate phase and n-butanol is more prominent.We choose the chromatographic separation technology combined with the DPPH free radical scavenging method to track and separate the active components in the ethyl acetate phase and the n-butanol phase.Structure of these compounds were identified by nuclear magnetic resonance data,and identified to be palmatine(1),3-O-?-d-glucopyranosyl-30-[32,33,34-trimethylpentyl]-16-ene-hopan(2),butyl-?-D-glucopyranosiden(3),n-butyl-?-D-fructopyranoside(4),beta-sitosterol(5),isovanillic acid(6),vanillin(7),and caroside(8).We examined the antioxidant activities of the obtained compounds,and found that all eight compounds had antioxidant activities,among which palmatine showed higher activity than that of the Vc positive control group.The cytotoxicity of compounds to SH-SY5Y cells was detected by MTT assay.The oxidative damage model of SH-SY5Y cells was established with H₂O₂,and the protective effect of compounds on SOD activity of oxidative damaged cells was explored.The results showed that palmatine is minimal toxic to SH-SY5Y cells at concentrationsbelow20?g/m L.3-O-?-d-glucopyranosyl-30-[32,33,34-trimethylpentyl]-16-ene-hopan is minimal toxic to SH-SY5Y cells at concentrations below 100?g/m L.n-butyl-?-D-fructopyranoside is non-toxic in SH-SY5Y cells at less than 20?g/m L.And when the concentration is lower than 50?g/m L,the SOD activity level of oxidative damaged cells after treatment with palmatine and n-butyl-?-D-fructopyranoside is significantly increased.This study revealed the great potential of the secondary metabolites of G.microrrhynchum as antioxidants and fungicides.At present,G.microrrhynchum has not been effectively developed and utilized,and our work will provide a valuable reference for subsequent related research.