An improved PCR method for direct identification of Porphyra(Bangiales,Rhodophyta) using conchocelis

来源 :Chinese Journal of Oceanology and Limnology | 被引量 : 0次 | 上传用户:wangxiao8910
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An improved method of PCR in which the small segment of conchocelis is amplified directly without DNA extraction was used to amplify a RUBISCO intergenic spacer DNA fragment from nine species of red algal genus Porphyra(Bangiales,Rhodophyta),including Porphyra yezoensis(Jiangsu,China),P.haitanensis(Fujian,China),P.oligospermatangia(Qingdao,China),P.katadai(Qingdao,China),P.tenera(Qingdao,China),P.suborboculata(Fujian,China),P.pseudolinearis(Kogendo,Korea),P.linearis(Devon,England),and P.fallax(Seattle,USA).Standard PCR and the method developed here were both conducted using primers specific for the RUBISCO spacer region,after which the two PCR products were sequenced.The sequencing data of the amplicons obtained using both methods were identical,suggesting that the improved PCR method was functional.These findings indicate that the method developed here may be useful for the rapid identification of species of Porphyra in a germplasm bank.In addition,a phylogenetic tree was constructed using the RUBISCO spacer and partial rbcS sequence,and the results were in concordant with possible alternative phylogenies based on traditional morphological taxonomic characteristics,indicating that the RUBISCO spacer is a useful region for phylogenetic studies. An improved method of PCR in which the small segment of conchocelis is amplified directly without DNA extraction was used to amplify a RUBISCO intergenic spacer DNA fragment from nine species of red algal genus Porphyra (Bangiales, Rhodophyta), including Porphyra yezoensis (Jiangsu, China) , P.haitanensis (Fujian, China), P.oligospermatangia (Qingdao, China), P.katadai (Qingdao, China), P.tenera (Qingdao, China), P. suborboculata Kogendo, Korea), P.linearis (Devon, England), and P. fallax (Seattle, USA). Standard PCR and the method developed here are both used using primers specific for the RUBISCO spacer region, after which the two PCR products were sequenced. The sequencing data of the amplicons obtained using both methods were functional, suggesting that the improved PCR method was functional. the findings of that method that developed the may be useful for the rapid identification of species of porphyra in a germplasm. , a phylogenetic tree was constructed using the RUBISCO spacer and partial rbcS sequence, and the results were in concordant with possible alternative phylogenies based on traditional morphological taxonomic characteristics, indicating that the RUBISCO spacer is a useful region for phylogenetic studies.
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