Study On The Biological Characteristics Of Symbiotic Bacteria Of Gastrodia Elata And The Changes Of Components Before And After Introduction Of Gastrodia Elata

Gastrodia elata B1 completes its own growth and development,must rely on its symbiotic bacteria(Fr.:Fr.)Staude to provide nutrition.Therefore,the quality of Armillaria directly affects the yield and quality of Gastrodia elata.The Armillaria genus in China is rich in germplasm resources,but there are few strains that can make Gastrodia elata stable and high-yield,and it is easy to degenerate.Therefore,the selection of dominant Armillaria strains and a medium suitable for the growth of Armillaria has become the focus of this topic research.The introduction of Gastrodia elata can expand the cultivation range,develop commodity production and give full play to the excellent characteristics of Gastrodia elata.However,whether the efficacy of Gastrodia elata after introduction is stable and how it has changed compared with the composition and content before introduction has become the main research direction of this article.Separation,purification,cultivation,comparison of morphological characteristics of Gastrodia elata symbiotic bacteria collected from different producing areas.At the same time,based on the rDNA-ITS sequence,12 purified strains were molecularly identified,and a total of 5 species of Armillaria were identified,named Armillaria mellea(Vahl.:Fr.)Kumm.?Armillaria gallica Marxm.&Romagn.?Armillaria cepistipes Velen.?Armillaria sinapina Berube.&Dessur.and Armillaria ostoya(Romagn.)Herink.And one non-Armillaria M2 Heydenia alpina.Terpenes and polysaccharides account for a large proportion in Armillaria mycelium.Based on the UV spectrophotometer and cultivation experiment method,discussion on the contents of total polysaccharides and total terpenes of 12 strains of Gastrodia elata symbiosis mycelium collected from 7 different origins.Results:The total terpenes content of Armillaria mycelium collected from Zhaotong,Yunnan was the highest,and the total terpenes content of Jinzhai mycelium was the lowest,and the total terpenes content fluctuated between 1.3%and 3.0%.The total polysaccharides content of Armillaria from Yichang,Hubei is the highest,and the content of polysaccharides from Armillaria in Diantan,Yunnan is the lowest,and the total polysaccharide content varies from 0.35%to 1.75%.The dry weight of mycelia was significantly positively correlated with glucose content and growth rate.Armillaria mellea A9 is commonly used in laboratory companion cultivation of Gastrodia elata,but in recent years,bacterial degradation and even inactivation often occurred.In this study,plate culture was adopted,and a more scientific and reasonable method was used to screen the optimal nutritional conditions(carbon source,nitrogen source,trace elements,vitamins)and environmental conditions(pH,temperature,light)for Armillaria mellea A9 growth.The results showed that the appropriate pH range of Armillaria mellea was 4.0?9.0 under semi-solid culture conditions,the temperature was about 25?,and the light condition was dark culture.Under the experimental conditions,the optimal carbon source is 25 g/L ethanol;the optimal nitrogen source is 2.5 g/L of soybean peptone;the optimal trace elements and vitamins are 0.5 g/L MnSO4,0.5 g/L MgSO4,and 20 mg/L vitamin B6.Experiments also show that adding natural carbon and nitrogen sources can promote the growth of Armillaria mellea mycelium.This study also provides a theoretical basis for the screening of Armillaria mellea rejuvenation medium.An ultra-high performance liquid chromatography(UPLC)method was established to simultaneously determine the contents of six main active ingredients before and after domestication of fresh Gastrodia elata from six habitats.The contents of six ingredients in Gastrodia elata samples from different producing areas are quite different,indicating that the place where Gastrodia elata is produced has a direct effect on the content of active ingredients in Gastrodia elata.The contents of gastrodin,parisonA,parisonB and parisonC in Gastrodia elata after introduction and domestication had obvious correlation,and the changes showed a certain trend.The microwave digestion method was used to process Gastrodia elata samples,and the inductively coupled plasma mass spectrometry(ICP-MS)method was used to simultaneously determine B?Na?Mg?Al?Si?P?K?Ca?Cr?Mn?Fe?Ni?Cu?Zn?As?Se?Mo?Cd?Hg?Pb content of 20 elements.The content of K in Gastrodia elata is much higher than other elements.The content of K in Gastrodia elata from the Northeast is even as high as 12.36 g/kg.Mg,Si,P,and Ca elements also account for a large proportion of Gastrodia elata,and Cu and Se elements are extremely small in Gastrodia elata,less than 5 mg/kg,the content of each element does not change significantly.After the introduction of Gastrodia elata from Northeast and Zhaotong to Tengchong,Yunnan.The content of Fe and Zn decreased and the content of Mn increased.