Identification The Origin Of Chinese Herb Guanzhong And Polypodiaceae By DNA Barcoding Technique

DNA barcoding,as one of useful and clipping species identification methods. Can be used as an important complement to traditional Chinese medicine (TCM)identification system, also an new tool to ensure clinical medicine accurate and effective. In this study, use of DNA barcoding to identification of the origin plant of Chinese herb Guanzhong and Polypodiaceae.ObjectiveFeasibility of DNA barcoding for species identification medicinal plants of Guanzhong herbs and Polypodiaceae two groups. In order to provide demonstration for TCM homonym phenomena and construction of DNA barcoding to identify medicinal ferns system, accumulation of information to improve the identification of medicinal plant DNA barcoding platform.MethodsIn our study,we priority screening test sequence Under bioinformatics prediction, early experimental basis and the current international consensus. 11 kinds of ferns representative chloroplast genome-wide data for multiple sequence alignment under progressive Mauve with bioinformatics prediction. And then collect locally collinear blocks (LCB), some LCB were f inded as candidate sequence universal primer, appropriate length and variability based on Primer、 Oligo et.DNAs were extracted from Polypodiaceae (38 samples 23 species) and Guanzhong (110 samples 67 species). Two candidate DNA barcodes of psbA-trnH and ITS2 were amplified and sequenced bidirectionally. Sequence quality check and contig generation were performed by using the CodonCode Aligner. Comparison of sequences obtained rate, inter- and intra-specific differences, species identification capability analysis.Results1) psbA-trnH obtained in all samples in medicinal plants Polypodiaceae(38 samples 23 species), the sequence length from 257 to 446bp, alignment length is 484bp and variable sites is 185 (38.2%). Alignment length and variable sites plus to 546bp and 266 (48.7%) in a wider range(134 samples 76 species).Based on psbA-trnH sequence of DNA barcoding technology to Polypodiaceae species identification analysis,the success rate was 82.6%(38samples of 23 species), Microsorum membranaceum and Microsorum superficiale are failure. Further integration Genbank data analysis (134 samples of 76 species), the success rate dropped to 61.8%, but congeneric species were clustered in the same branch with N-J.2) For Guanzhong study,110 samples 67 species sequenced successful with psbA-trnH. The sequence length from 360 to 620bp, alignment length is 638bp and variable sites is 485bp (76.0%). ITS2 obtained rate is 81.6% of 36 samples 21 species in Guanzhong. Sequence length 154bp-327bp, alignment length 391bp and the variable sites persent 361 (92.3%)In the 67 species of Guanzhong,48 are effectively identified by psbA-trnH with a identification success rate 71.6%. In addition, ITS2 could identify the other psbA-trnH identification failure or inter- and intra- too large’ species successfully. While for ITS2 sequence, the discriminative efficiency iecreased with more within-species variation larger than between-species ones in a wider range.When combined psbA-trnH and ITS2 sequences, the overall efficiency of the species identified 98.5%(66/67). Dryopteris erythrosora was identification of failure.ConclusionpsbA-trnH could serve as the valuable reference to discriminate the more homonyms of guanzhong, and another assistant marker(e. g. ITS2) should be selected in future to improve the success rate in some genus. And psbA-trnH sequence could be used as medicinal plants Polypodiaceae recommended sequence for DNA barcoding. But also need to clearly identify the extent, while discriminative efficiency decreased in a wider range.