The Study Of The Quality Of The Ginseng And The Biodiversity Of Endophytic Bacteria About Seeds

In order to evaluate quality of panax ginseng from TCM material market,we collect 60 panax ginseng samples from AnHui BoZhou TCM material market and Harbin Sankeshu TCM material market.We carry out the research on the contents of ginseng saponins,flavonoids and polysaccharide in panax ginseng samples for quality evaluation.Before the study,the samples were pulverized and sieved by 20 mesh screen for expanding the contact area between the sample and the extraction solvent,in order to improve the extraction efficiency better.Polysaccharide in panax ginseng materials were extracted by water,Polysaccharide is a kind of natural ingredients with strong adhesion,so it is diffcult to extract polysaccharides when the particle size of sample is small.After the extraction,the water extracts were filtrated by gauze and then centrifuged to get rid of water,Afterwards,sevage reagents were added into the extracts for deprotein,and centrifuged to recover the supernatant.Ethanol was added into the supernatant,and the precipitation obtained is ginseng polysaccharide.Ginseng polysaccharide extraction process was optimized by the single factor and orthogonal experiments（L₉（3⁴））,and polysaccharide was detected by the phenol-sulfuric acid method Flavonoid is also a kind of important natural ingredients in ginseng panax,ethanol solution was used to extract flavonoids,and the extraction process was optimized by the orthogonal experiment（L₉（3⁴））.According to Chinese Pharmacopoeia,HPLC-PDA was used to determine the content of ginseng saponins（Rg,Re,and Rbl）at the wavelength of 204 nm.The quality of ginseng was evaluated by detecting the contents of ginseng saponins（Rg,Re,and Rb1）.Meanwhile,the residual organochlorine pesticides（DDT、BHC and PCNB）in panax ginseng were detected by the GC method according to Chinese Pharmacopoeia.Finally,454 sequencing technique was used to detect the endophyte diversity of 5 panax ginseng seed samples.1.After the single factor investigation,the orthogonal experiment（L₉（3⁴））was used to optimize the extraction process of polysaccharides.Results showed that temperature（A）>time（B）>extraction times（C）>error（D）,and the optimal extraction parameters is extraction temperature 100℃,extract time 4h,extraction cycle 2 times,and liquid to solid ratio 1:20.Ethanol sinking process is optimized by the single factor investigation,that the optimal extraction parameters is ethanol concentration 85%,and ethanol time 24h.Polysaccharides were extracted from 60 panax ginseng samples collected in different TCM material markets using the above methods and the content of polysaccharides range from 10%to 20%,and the average content is 15.4%.2.After the single factor investigation,the orthogonal experiment（L₉（3⁴））was used to optimize the extraction process of flavonoids.Results showed that the order of the effect of parameters on the extraction efficiency is alcohol（A）>extraction time（C）>the volume of solvent（B）>error（D）,and the optimal extraction parameters is alcohol concentration 85%,extraction time 3h,the ratio of liquid to solid material 15,and extraction cycle 3 times.Flavonoids were extracted from panax ginseng samples using the above methods,and the total flavonoid contents are determined by the ultra violet-visible spectrophotometry method,and the content of polysaccharides range from 11.35 mg to 27.35 mg,and the average content is 0.9493 mg/g.3.According to Chinese Pharmacopoeia,HPLC-PDA was used to measure the content of ginseng saponins at the wavelength of 204 nm.We founded that the ginseng saponin contents（Rg1+Re）in the samples（10,18,31,3⁴,48,59）are lower than the level reported by Chinese Pharmacopoeia;the ginseng saponin（Rbl）content in samples（4,6,9,39,55,57）is lower than 0.2%.The quality fraction defective of samples is 20%.Compared with the ginseng saponins content in samples collected from different origin places.We discover that the quality of panax ginseng samples collected from Jilin is the best,.LiaoNing is the second,and HeilongJiang is relatively the poorest.4.We use the GC method to measure the content of the residual pesticide（DDT,BHC,and PCNB）in panax ginseng samples.We found that the PCNB in the samples（2,18,and35）surpass 0.1mg/kg;the DDT in the sample（4）surpass 0.2g/kg;the BHC in sample（6 and 56）surpass 0.2mg/kg.The average contents of BHC,DDT and PCNB are to 97.96,58.95,46.87.According to Chinese Pharmacopoeia,the samples（2,4,6,9,10,18,31,35,36,48,56 and 59）are not fitted for the provisions,making up about 20%in total.5.The regression equation was forecasted using SPSS:Y=0.017X+0.009Z+0.356,R=0.588（X is the flavonoids content,units:mg/20g;Y is the polysaccharide content,units:g/5g;Z is ginseng saponin content（Rg1+Re+Rb1））.It can be concluded from the regression equation,that polysaccharide content is positively correlated with those of the ginseng saponinsand flavonoids;polysaccharide content was set as independent variable（X）,and flavonoids content was set as dependent variable（Y）,A regression equation was established as Y=20.379X+4.071,R=0.584,Based on the equation,polysaccharides are positively correlated with flavonoids.Polysaccharide content was set as independent variable（X）,and ginseng saponin content was set as dependent variable（Y）.A quadratic regression equation was established as Y=-24.621X2+37.859X-7.783,R=0.532>0.5.Based on the equation,polysaccharides are positively correlated with flavonoids.When the content of polysaccharides is 15.7%,the ginseng saponin is maximum.（When X<0.7688,it is positive,or negative）.6.454 sequencing technique was used to detect the endophyte diversity in 5 panax ginseng seed samples.Results showed that Cyanobacteria,proteusbacillus vulgaris are dominant fungi.Besides,Bacteroides and Actinomycete are also detected.The species in panax ginseng is relatively simple,which might be the reason that panax ginseng seedlings are prone to disease.