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WU Chunhui, JIANG Peng, ZHAO Jin, FU Huihui. High efficiency of protoplast preparation for artificially cultured Ulva prolifera (Ulvophyceae, Chlorophyta)[J]. Journal of Oceanology and Limnology, 2018, 36(5): 1806-1811

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

WU Chunhui1,2,3, JIANG Peng1,2, ZHAO Jin1,2, FU Huihui1,2
1 Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China;
2 Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China;
3 University of Chinese Academy of Sciences, Beijing 100049, China
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)×106 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.
Key words:    protoplast|Ulva prolifera|regeneration|artificial culture   
Received: 2017-03-01   Revised:
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