Fingerprints Study On Ferula Ferulaeoides(Steud.) Korov. From National Medicine In Xinjiang

Objective:To establish GC-MS fingerprints of essential oils and DNA molecule fingerprints of Ferula ferulaeoides for assessment and quality control of Ferula ferulaeoides. Methods:(1) The essential oils of8habitats from F. ferulaeoides were extracted by steam distillation method and their chemical constituents were analyzed by GC/MS.(2) Genomic DNA was extracted by three different methods.(3) The ISSR-PCR amplification system on F. ferulaeoides in concentrations of MgCl2, dNTPs, primers, Taq polymerase and template DNA were optimized by single factor test and orthogonal experimental design test, and the PCR result was analyzed by SPSS.(4) Using orthogonal design test combined with the single factor gradient optimization method, the geranium ISSR-PCR system of F. ferulaenides was determined. To study the genetic diversity and cluster analysis of8wild populations.(5) By comparing the results of different amplification, and ultimately selected15ISSR primers and15RAPD primers were used to build F. ferulaenides fingerprints. Results:(1) The method on GC-MS fingerprints of essential oils of F. ferulaeoides was established and the GC-MS fingerprints of essential oils of F.ferulaeoides, showing12characteristic peaks.44samples were classified into two group.(2) Genomic DNA Kit method was higher than the improved CTAB method and improved SDS method with more quality, more clear bands, shorter time and more simple operation.(3) ISSR:Most of the factors in different levels had the significant effects on the result of PCR, and the most remarkable factor was the concentration of Mg2+, dNTPs and primers. RAPD:The most remarkable factor was the concentration of Mg2+, dNTPs and primers.(4) ISSR:Fifteen primers were selected to produce highly reproducible ISSR bands. Among126amplified bands,93showed polymorphism, the percentage of polymorphic bands (PPB)reached73.8%. RAPD:Fifteen primers were selected to produce highly reproducible RAPD bands. Among226amplified bands,203showed polymorphism, the percentage of polymorphic bands(PPB) reached89.8%.(5) Calibration the ISSR and RAPD characteristics of DNA bands. Conclusion:(1) The method of GC-MS fingerprinting was simple, accurate with good reproducibility and can be used for the comprehensive quality evaluation of Ferula ferulaeoides.(2) Genomic DNA Kit method was the optimal method for plant medicines belongs to the genus ferula and can be used directly in downstream molecular biology experiments.(3) The established and optimized ISSR and RAPD reaction system is stable and credible according to the testing results of22samples of F. ferulaeoides, and provides the basis for further study on identification and genetic diversity analysis of Ferula ferulaeoides by ISSR molecular marker technique.(4) F.ferulaenides holds high genetic diversity and the majority of genetic variation occurs among the populations.(5) Using DNA fingerprinting can be fast, accurate identification in Ferula species. To ensure ferula source in accurate classification is meaningful.(6) F.ferulaenides from different areas have a certain correlation between the oil composition changes and genetic variation.