Study The Quality Evaluation Of Paris Polyphylla Var.Yunnanensis Transforming From Wild Into Cultivated Variety And Its Rhizosphere Soil Bacteria

ObjectiveTo study the quality and the difference about main effective components on transforming wild Paris polyphylla var.yunnanensis into cultivated variety in Yunlong,Fengyi,Nanjian,Jianchuan et al.and preliminary study the correlation between the quality and bacteria from rhizosphere of Paris polyphylla var.yunnanensis in Fengyi Dongshan of Dali,Tuanjie Dadeng of Yunlong,Qingsong of Yunlong and Yousuo Qisheng of Eryuan.MethodsAnalyze the quality and the difference about main effective components by HPLC and UPLC-ESI-Q-TOF-MS,isolate and purify by plate dilution coating method and streak separation method,and identify based on the 16 S r DNA sequences.Results1.There are significant(P<0.05)or highly significant(P<0.01)differences in Polyphyllins I,II,V,VI,VII,H and Dioscin in the rhizomes of transforming wild Paris polyphylla var.yunnanensis into cultivated variety from 18 plant bases.The total contents of Polyphyllins I,II,VI,VII(0.4594-2.9674% mg/g)in the rhizomes of Paris polyphylla var.yunnanensis is lower than the 0.6% limit specified in the 2015 edition of the Pharmacopoeia of the People's Republic of China except that in the rhizomes of Paris polyphylla var.yunnanensis from Madeng of Jianchuan,Leqiu of Nanjian and Guanping of Yunlong.A total of 22 peaks of HPLC fingerprint of the rhizomes of Paris polyphylla var.yunnanensis in different plantings were common,the similarity of HPLC fingerprint was higher(0.907-0.988),the RSD of relative retention time(0.06%-2.55%)of each total peak was smaller but the relative peak area(3.80%-105.90%)of each total peak was larger.2.The steroid components in the rhizomes and leaves of Paris polyphylla var.yunnanensis in 16 different bases were detected for qualitative analysis by UPLC-MS in positive ion mode,and then 13 steroid components(Polyphyllin I,II,V,VI,VII,D,H,G,Pseudoproto-pb,Pariyunnanoside B,Dioscin,Gracillin,Tg)were totally identified by it.According to PCA mode,the principal components in the rhizomes of Paris polyphylla var.yunnanensis can be roughly clustered into 2 categories,the time of the major differences peak were 0.53 min(m/z 130.500)and 6.53 min(m/z 366.500),and meanwhile the principalcomponents in the leaves can be roughly well clustered,the time of the major differences peak were 45.53 min(m/z 398.500),36.53 min(m/z 396.500,412.500)and 6.53 min(m/z366.500).3.A total of 74 bacterial strains were obtained from rhizosphere soil of Paris polyphylla var.yunnanensis in 4 plant bases,including 23 actinobacteria strains and 51non-actinobacteria strains,2 actinobacteria strains and 9 non-actinobacteria strains(Fengyi Dongsshan of Dali),6 non-actinobacteria strainssix Pantoea strains(Tuanjie Dadeng of Yunlong),20 actinobacteria strains and 29 non-actinobacteria strains(Qingsong of Yunlong),1 actinobacteria strains and 7 non-actinobacteria strains(Yousuo Qisheng of Eryuan).There are significant(P<0.05)or highly significant(P<0.01)differences in Polyphyllins I,II,V,VI,VII,H and Dioscin in the rhizomes of Paris polyphylla var.yunnanensis into cultivated variety from 4 plant bases.Among these Paris polyphylla var.yunnanensis into cultivated variety of 4 plant bases,the total content of Polyphyllins I,II,VI and VII in the rhizomes of Paris polyphylla var.yunnanensis in Tuanjie Dadeng of Yunlong(1.9975% mg/g)was highest,and that in Yousuo Qisheng of Eryuan(1.9892% mg/g)was the second.Conclusion1.There are highly significant(P<0.01)or significant(P<0.05)differences in Polyphyllins I,II,V,VI,VII,H and Dioscin in the rhizomes of 18 transforming wild Paris polyphylla var.yunnanensis into cultivated variety from the above-mentioned 18 plant bases,the similarity of HPLC fingerprint is higher and the quality is good.2.The distribution differences of 13 main effective components,including Pseudoproto-pb,Polyphyllin G,Pariyunnanoside B,Polyphyllin VII,Polyphyllin D,Polyphyllin H,Polyphyllin VI,Polyphyllin II,Dioscin,Gracillin,Polyphyllin I,Tg and Polyphyllin V)of steroid saponins in the rhizomes and leaves of Paris polyphylla var.yunnanensis in 16 different plantings are greatly different.Meanwhile,the differences of principal components in the rhizomes are higher but smaller in the leaves based on the PCA mode.3.We find that the strains of the genera Micromonospora,Enterobacter,Pantoea and Stenotrophomonas maltophilia strains were related to the quality of transforming wild Parispolyphylla var.yunnanensis into cultivated variety,which maybe could promote the contents of effective components of Paris polyphylla var.yunnanensis.