Identification of plants in Fritillariae L. by DNA barcoding technology
10.7501/j.issn.0253-2670.2014.11.021
- Author:
Chao YU
1
Author Information
1. Institute of Biotechnology, Zhejiang Wanli University
- Publication Type:Journal Article
- Keywords:
DNA barcoding;
Fritillaria L.;
Identification;
ITS1;
ITS2;
Molecular
- From:
Chinese Traditional and Herbal Drugs
2014;45(11):1613-1619
- CountryChina
- Language:Chinese
-
Abstract:
Objective: DNA barcoding technology was employed to compare the identification efficiency of ITS1and ITS2 sequences in plants of Fritillariae L. Methods: Twenty-one samples from eight species of Fritillariae L. were collected. Genomic DNAs from these samples were extracted, ITS1 and ITS2 sequences were amplified by PCR, and the PCR products were sequenced. The DNA extraction rate, PCR amplification rate, and sequencing achievement ration were compared. The identification efficiency of two sequences was evaluated by BLAST-Search, the distribution of genetic distance was analyzed by SPSS software, and the phylogenetic trees were constructed by MEGA software. Results: Comparing the primary structures of ITS1 and ITS2 sequences amplified from different species, the results showed that the sequence length of IT1 was 205-229, informative sites were 27 and occupied 11.8%, and the identification achievement rate was 72.2%; The sequence length of IT2 was 236-244, informative sites were 20 and occupied 8.2%, and the identification achievement rate was 100%. Intra-specific and inter-specific genetic distances of ITS1 were 0.001 8 and 0.066 1, and those of ITS2 were 0.000 5 and 0.046 8, respectively; The results also showed that ITS2 was relatively conservative in intra-species; The NJ phylogenetic trees showed that these two sequences could accurately identify Fritillaria thunbergii, F. cirrhosa, and north-Fritillaria, but phylogenetic trees based on ITS2 had a higher degree of accuracy. Conclusion: ITS2 sequence is optimal as barcoding sequence for Fritillaria L. genus and is more suitable for identifying the different species of Fritillaria L.
