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High efficiency of protoplast preparation for artificially cultured Ulva prolifera (Ulvophyceae, Chlorophyta)

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Abstract

Protoplast isolation was relevant for gene manipulation in Ulva, and universal protocols have been proposed based on evaluation for various wildly collected species. However, only clonal laboratory cultures were practical for genetic transformation, and whether applicability of such universal protocol existed for these artificial cultures has never been investigated. In this research, samples in different physiological states or developmental stages were tested in U. prolifera. The results proved that the protoplast yields were strongly dependent on the characteristics of samples. Neither Fv/Fm value nor chlorophyll content exhibited an ideal correlation with the protoplast yields. Alternatively, specific growth rate, coupled with developmental stage, could serve as an effective combined index to determine the right time for protoplast isolation. According to this instruction, here we reported the highest yields of protoplast ((31.5±1.9)×10 6 cells/g f. wt.) in U. prolifera, following comparison between protocols, and further optimizations on enzyme content, incubation period, starting biomass and pretreatment. This specified protocol for artificially cultured clonal samples could meet the need for protoplast-mediated genetic transformation in U. prolifera.

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Authors and Affiliations

  1. Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China

    Chunhui Wu  (吴春辉), Peng Jiang  (姜鹏), Jin Zhao  (赵瑾) & Huihui Fu  (付慧慧)

  2. Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266237, China

    Chunhui Wu  (吴春辉), Peng Jiang  (姜鹏), Jin Zhao  (赵瑾) & Huihui Fu  (付慧慧)

  3. University of Chinese Academy of Sciences, Beijing, 100049, China

    Chunhui Wu  (吴春辉)

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  1. Chunhui Wu  (吴春辉)
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  2. Peng Jiang  (姜鹏)
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  3. Jin Zhao  (赵瑾)
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  4. Huihui Fu  (付慧慧)
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Corresponding author

Correspondence to Peng Jiang  (姜鹏).

Additional information

Supported by the National Natural Science Foundation of China (No. 41776153), the Scientific and Technological Innovation Project financially supported by Qingdao National Laboratory for Marine Science and Technology (No. 2016ASKJ02-1), the Strategic Priority Research Program of the Chinese Academy of Sciences (No. XDA11020304), and the Postdoctoral Application Research Program funded by Qingdao (No. 2016189)

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Wu, C., Jiang, P., Zhao, J. et al. High efficiency of protoplast preparation for artificially cultured Ulva prolifera (Ulvophyceae, Chlorophyta). J. Ocean. Limnol. 36, 1806–1811 (2018). https://doi.org/10.1007/s00343-018-7058-0

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  • DOI: https://doi.org/10.1007/s00343-018-7058-0

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