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 Porphy...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.展开更多
Cryopreservation of Porphyra yezoensis conchocelis was conducted with cryoprotectants and a proposed pretreatment procedure and thawing methods explored. Six cryoprotectants combined by DMSO with ethylene glycol(EG),p...Cryopreservation of Porphyra yezoensis conchocelis was conducted with cryoprotectants and a proposed pretreatment procedure and thawing methods explored. Six cryoprotectants combined by DMSO with ethylene glycol(EG),propylene glycol(PEG),sorbitol and sucrose were developed. The effect of prefreezing at - 40℃ or -20℃ for different time durations was compared and the thawing methods were screened. It was shown that the cryoprotectant including 10% DMSO with 0.5 molL-1 sorbitol exhib-ited the optimal effect. The ideal pretreatment was that conchocelis segments were stayed for 20 min at - 40℃ before stored in liquid nitrogen,and 40℃ water bath was proper for quick thawing. The highest recovery rate of cryopreserved P. yezoensis conchocelis reached 89.41%.展开更多
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)+2 种基金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)
文摘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.
文摘Cryopreservation of Porphyra yezoensis conchocelis was conducted with cryoprotectants and a proposed pretreatment procedure and thawing methods explored. Six cryoprotectants combined by DMSO with ethylene glycol(EG),propylene glycol(PEG),sorbitol and sucrose were developed. The effect of prefreezing at - 40℃ or -20℃ for different time durations was compared and the thawing methods were screened. It was shown that the cryoprotectant including 10% DMSO with 0.5 molL-1 sorbitol exhib-ited the optimal effect. The ideal pretreatment was that conchocelis segments were stayed for 20 min at - 40℃ before stored in liquid nitrogen,and 40℃ water bath was proper for quick thawing. The highest recovery rate of cryopreserved P. yezoensis conchocelis reached 89.41%.
