Separation And Identification Of Endophytic Fungi From Yingjing Gastrodia Elata

In this study,we have researched the endophytic fungi of Yingjing Gastrodia elata which has been planted a long history and has good quality,known as "Gong Ma".Endophytic fungi community structure of Yingjing Gastrodia elata and part of the chemical composition have been studied,and a strain of endophytic fungi has been screened out,which has good antioxidant activity and can produces Gastrodine.This study was laid the theoretical basis for the development and utilization of endophytic fungi of Yingjing Gastrodia elata.The main results were as follows:1?Analysis of endophytic fungi community from Yingjing Gastrodia elataUsing the high-throughput sequencing and comparing to NCBI databasest,we analyze the endophytic fungi community from Yingjing Gastrodia elata,compared with Baoxing Gastrodia elata.Endophytic fungi community shannon index of Yingjing Gastrodia elata and Baoxing Gastrodia elata respectively were 2.6889 and 2.6889,which showed that endophytic fungi community of Yingjing Gastrodia elata has better diversity than Baoxing Gastrodia elata.Yingjing Gastrodia elata endophytic fungi community has good species diversity and they can divided into one branche in the clustering analysis.There is a strong uniqueness with species diversity of Yingjing Gastrodia elata endophytic fungi.2?Separation and identification of endophytic fungi from Yingjing Gastrodia elataUsing the Gastrodia elata from Siping where the main production area of Yingjing as experimental materials,we isolated 15 strains endophytic fungi in the PDA medium,including 3 strains of Fusarium sp,5 strains of Neonectria sp,5 strains of Ascomycota sp,2 strains of Oidiodendron sp.and 1 strains of Dothiorda sp.Other 4 strains of fungi have not clear attribution,which may be rarely studied.3?Determination of total phenols and total flavonoid content of endophytic fungi from Yingjing Gastrodia elataTaking gallic acid as standand,total phenolic contents from endophytic fungi were respectively measured by Folin-Ciocalteu's colorimetric method and taking rutin as standand,total flavonoid content was measured by colorimetric method.The total phenols contents and total flavonoid contents of the 10 endophytic fungi were investigated.The total phenols contents varied from 5.37 to 9.68 mg/g of dry weight.The highest concentration of phenols was found in the extract of GeR3(9.68±0.41 mg/g),followed by GeR6(9.22±0.09 mg/g).The total flavonoid contents varied from 1.85 to 6.68 mg/g of dry weight.The highest concentration of flavonoid was observed in extract of GeR7(6.68±0.19 mg/g)followed by GeR3(5.52±0.23 mg/g).4.Gastrodine content in the endophytic fungi of Yingjing Gastrodia elataWith reference to the Chinese pharmacopoeia(2015 edition),the gastrodine content of the 10 strains of endophytic fungi mycelium were determined using HPLC,Acetonitrile-water(3:97)is used as mobile phase.After the examination,GeR6(Oidiodendron sp,)gastrodine content in dry mycelium was 39.2 ?g/g.It provides a new source of gastrodine and is conducive to the protection of the Gastrodia elata wild resources.5.Determination of antioxidant activity of mycelia ethanol extractsTaking butylated hydroxytoluene(BHT)as positive control,,the antioxidant activity of mycelia ethanol extracts of 10 strains endophytic fungi were detected.The T-AOC value was in the range of 367.61 to 3186.66 U/g,among which,the value of GeR6(3186.66 U/g)was significantly higher than others,but lower than BHT.In the DPPH experiments,the GeR6(IC50=456.42 ?g/mL)also showed good antioxidant activity,which suggest that it could be a promising resource of natural antioxidants.