An improved PCR method for direct identification of Porphyra(Bangiales,Rhodophyta) using conchocelis based on a RUBISCO intergenic spacer  

作　　者：王超 董栋 王广策 张宝玉 彭光 许璞 汤晓荣 

机构地区：[1]Key Laboratory of Experimental Marine Biology,Institute of Oceanology,Chinese Academy of Sciences [2]Graduate University of Chinese Academy of Sciences [3]Tianjin University of Science and Technology [4]Changshu Institute of Technology [5]Ocean University of China

出　　处：《Chinese Journal of Oceanology and Limnology》2009年第3期513-518,共6页中国海洋湖沼学报（英文版）

基　　金：Supported by the National High Technology Research and Development Program of China (863 Program)(No 2006AA10A402);Project for Supporting National Development (No 2006BAD09A04);the National Natural Science Foundation of China (Nos U0633006,40476059);the Natural Science Foundation of Qingdao (No 05-2-p-2);the Knowledge Innovation Program of the Chinese Academy of Sciences (No KZCX2-211)

摘　　要：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 （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.

关 键 词：RHODOPHYTA PORPHYRA RUBISCO intergenic spacer DNA sequence PCR 

分 类 号：S917.3[农业科学—水产科学]