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Rapid DNA extraction from conchocelis and ITS-1 rDNA sequences of seven strains of cultivated Porphyra yezoensis (Bangiales, Rhodophyta)

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Abstract

In order to extract DNA rapidly from cultivated Porphyra, we extracted total DNA from conchocelis using the ISOPLANT II kit (Nippon Gene) without liquid nitrogen treatment or CsCl-gradient ultracentrifugation. By confirming the reproducibility of RAPD patterns, it is concluded that the quality of the extracted DNA is sufficient to use as a template for molecular investigation. Using this rapid method, the nuclear ribosomal DNA of the internal transcribed spacer (ITS) regions was amplified from seven strains of cultivated Porphyra, which had been maintained as free-living conchocelis by subculturing in the laboratory. From the amplified DNAs, the ITS-1 sequences were determined in order to identify the species and genetic relationship of the strains. The sequences were identical in the seven strains, and all the strains were identified as P. yezoensis. Furthermore, the gametophytic blades of these strains showed long linear or oblanceolate shapes in the laboratory culture. It was concluded that these strains are P. yezoensis form. narawaensis. This rapid DNA extraction method from conchocelis will be a powerful tool for phylogenetic analysis and for genetic improvement of cultivated Porphyra.

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Authors and Affiliations

  1. Fisheries Technology Institute, Hyogo Prefectural Technology Center for Agriculture, Forestry & Fisheries, Akashi, 674-0093, Japan

    Kyosuke Niwa

  2. Department of International Agriculture Development, Tokyo University of Agriculture, Setagayaku, 156-8502, Japan

    Yusho Aruga

Authors
  1. Kyosuke Niwa
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  2. Yusho Aruga
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Niwa, K., Aruga, Y. Rapid DNA extraction from conchocelis and ITS-1 rDNA sequences of seven strains of cultivated Porphyra yezoensis (Bangiales, Rhodophyta). Journal of Applied Phycology 15, 29–35 (2003). https://doi.org/10.1023/A:1022998823920

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