| 著者 |
Shigekatsu Suzuki, Masakazu Hoshino, Kayoko Yamamoto, Shuhei Ota, Mary-Hélène Noël, Yuuhiko Tanabe, Shinya Uwai, Masanobu KAWACHI
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| Abstract |
The lack of broadly applicable axenization methods has fundamentally limited research across algae and protists, particularly for non-model species with complex morphologies and extensive microbial associations. Existing approaches typically require species-specific optimization, rely on antibiotics, or fail to remove tightly associated bacteria, preventing scalable and reproducible establishment of axenic cultures. Here, we present a broadly applicable, optimization-free framework for axenization based on two complementary methods: Cell Surface Sterilization (CSS) and Layered Gel Aseptic (LGA). Without antibiotics, we demonstrate successful axenization and maintenance across more than 80 strains spanning major algal and protist lineages, including species with complex colonies, extracellular polysaccharides, and macroalgal tissues. Furthermore, we show that this framework enables direct establishment of axenic strains from natural environmental samples, enhances single-cell genomics from xenic material, and supports semi-automated workflows when combined with cell sorting. These results establish axenization as a broadly solvable and scalable process, and provide an enabling framework for expanding experimental, genomic, and ecological studies beyond established model algal and protist systems.
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