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

doi:10.1007/s00343-009-9169-0

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	An improved PCR method for direct identification of Porphyra (Bangiales, Rhodophyta) using conchocelis based on a RUBISCO intergenic spacer
	Wang Chao 1,2; Dong Dong 1,2; Wang Guangce 1,3; Zhang Baoyu 1; Peng Guang 1; Xu Pu 4; Tang Xiaorong 5
刊名	CHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY
	2009-08-01
卷号	27 期号:3 页码:513-518
关键词	Rhodophyta Porphyra Rubisco Intergenic Spacer Dna Sequence Pcr
ISSN号	0254-4059
DOI	10.1007/s00343-009-9169-0
文献子类	Article
英文摘要	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.
学科主题	Limnology ; Oceanography
URL标识	查看原文
语种	英语
WOS记录号	WOS:000270742000012
公开日期	2010-12-22
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/3027]
专题	海洋研究所_实验海洋生物学重点实验室
作者单位	1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China 3.Tianjin Univ Sci & Technol, Tianjin 300222, Peoples R China 4.Changshu Inst Technol, Changshu, Peoples R China 5.Ocean Univ China, Qingdao 266071, Peoples R China
推荐引用方式 GB/T 7714	Wang Chao,Dong Dong,Wang Guangce,et al. An improved PCR method for direct identification of Porphyra (Bangiales, Rhodophyta) using conchocelis based on a RUBISCO intergenic spacer[J]. CHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY,2009,27(3):513-518.
APA	Wang Chao.,Dong Dong.,Wang Guangce.,Zhang Baoyu.,Peng Guang.,...&Tang Xiaorong.(2009).An improved PCR method for direct identification of Porphyra (Bangiales, Rhodophyta) using conchocelis based on a RUBISCO intergenic spacer.CHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY,27(3),513-518.
MLA	Wang Chao,et al."An improved PCR method for direct identification of Porphyra (Bangiales, Rhodophyta) using conchocelis based on a RUBISCO intergenic spacer".CHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY 27.3(2009):513-518.