| The root of Codonopsis pilosula(Franch.)Nannf.is the main source of a traditional Chinese medicine,Codonopsis Radix.Modern pharmacological studies have indicated that Codonopsis pilosula contains alkaloids,alkynes,terpenes,flavonoids and sugars,and it has a variety of biological activities,such as improving Gastrointestinal motility,anti-inflammatory,anti-oxidation.Therefore,Codonopsis pilosula is widely used in clinical treatment and prevention,in addition,it has also been utilized as a raw material for the development of various functional foods.With the uninterrupted expansion of the market and the increase in usage,the wild resources of medicinal plants are gradually exhausted.The current Codonopsis Radix is mainly derived from large-scale artificial cultivation.On the one hand,the use of PGRs will change the content of active ingredients in medicinal materials and affect the quality and efficacy of medicinal materials.In order to increase yield,improve quality and maximize economic benefits,plant growth regulator(PGR)has been used more and more frequently in the cultivation of Codonopsis pilosula in recent years.However,long-term uncontrolled abuse and excessive use make PGRs remain in medicinal materials and the surrounding environment,seriously threatening human health and drug safety.On the other hand,the impact of PGRs on the soil ecological environment cannot be ignored.The community structure and composition of soil microorganisms are important indicators for assessing the safety of PGRs to soil ecology.However,among various national and local regulations,the safety control of the use of PGRs for Codonopsis pilosula is not yet consummate,there is still a lack of convincing control indicators.Based on this,a comprehensive and systematic study of the metabolite spectrum of Codonopsis pilosula and the composition of the soil microbial community structure,and screening out specific PGR markers are of great significance for the safety assurance and the quality control of medicinal materials,and provide a scientific basis for evaluating the environmental safety of PGRs,establishing scientific and reasonable usage specifications,and later soil remediation.1.Reveal the regulation of chlormequat chloride on the metabolites of Codonopsis pilosula by Non-targeted metabolomics analysisMetabolomics of Codonopsis pilosula after application of chlormequat chloride(CC)was analyzed using ultrahigh performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry technology(UPLC-QTOF-MS).Compared with that of no application,the weight of fresh and dried roots treated by CC increased by 24.2%～63.8% and 25.0%～71.0% respectively,and the contents of total soluble sugars,total flavonoids and total saponins decreased by 10.4%～18.5%,32.4%～42.9%,26.3%～65.2%,respectively.The residue of CC in Codonopsis pilosula roots increased with a dose-dependent manner,which was 27.31～43.94-times than that of non-applying.Furthermore,the application of CC affected the structure and composition of the overall metabolites in the Codonopsis pilosula roots.In order to investigate the differences,21 potential markers were screened between samples treated by CC and without application based on univariate analysis combined with multivariate statistical analysis.Among them,11 metabolites such as atractylenolide III were down-regulated,while the other 10 metabolites,including jasmonic acid,were up-regulated after CC treatment.The results confirmed that application of CC promoted root growth and regulated metabolite profile of Codonopsis pilosula in a dose-dependent manner.This study also provides reliable candidates for discriminative chemical markers that can be applied to assess the quality of medicinal materials with and without PGRs.This study aims to reveal the influence of PGRs on the metabolite profile of Codonopsis pilosula and to obtain specific chemical markers.The results obtained will help to provide important information for the establishment of safety control indicators for the use of PGRs.2.The effect of chlormequat chloride on the microbial diversity in the soil cultivated with Codonopsis pilosula.16S rRNA and ITS gene library construction and sequencing were performed on the soil where Codonopsis pilosula was grown,and annotations at various taxonomic levels were completed.Comprehensive analysis and comparison of the soil microbial diversity and community structure composition with or without CC applied.The results of Alpha analysis showed that the application of CC reduced soil microbial diversity and affected species composition,and this effect seemed to be related to the frequency of application.Principal component analysis(PCA)proved that there were differences between the soil samples that between CC-sprayed and non-sprayed,while LEf Se analysis screened out the bacterial flora that caused significant differences.There were 155,202,and 162 different species between soil samples non-sprayed and each CC-sprayed(1 time,2 times and 3 times).Finally,the potential biomarkers related to CC application in the soil where Codonopsis pilosula were grown were determined,the bacteria include Micrococcales,Micromonosporaceae,Solirubrobacteraceae,Nocardioidaceae,Armatimonadetes,Chloroflexia,Deinococci,Proteobacteria,clostridium;the fungi include Dothideomycetes,Sordariomycetes,Agaricomycetes,Geminibasidiomycetes.The research results provide important information for assessing the risk of PGRs residues in the environment.Finally,the pearson correlation analysis between the significant difference metabolites and the difference flora was carried out,which showed that the microbial flora and multiple metabolites have different degrees of correlation.It shows that the reason why the application of CC can significantly change the abundance of metabolites in the roots of Codonopsis pilosula may be related to the change of the microbial community in the planting soil. |
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