Separation of cultured strawberry cells producing anthocyanins in aqueous two-phase system.
A rapid and simple selection method of high-yield cells has been desired to establish highly productive cell lines for useful secondary metabolites. For this purpose, a new attempt was made to partition cultured plant cells in a poly(ethylene glycol)-dextran aqueous two-phase system (ATPS). The applicability of the ATPS in partitioning cultured strawberry cells (designated FAW) was investigated. The result of single-step partitioning in the ATPS supplemented with 0.4 mmol/kg lithium sulfate showed that FAW cells cultivated for 7 d under light-irradiation were separated into two cell populations with significantly different anthocyanin content. Additionally, the analysis technique of microscopic cell images showed that cells accumulating a high level of anthocyanin were partitioned completely into the bottom phase in a partitioning experiment of FAW cells cultivated for 10 d under light-irradiation in the ATPS supplemented with 1.8 mmol/kg potassium phosphate buffer. These results indicated that cell partitioning in ATPS increased the intracellular anthocyanin content and that the cultured strawberry cell population was heterogeneous in terms of cell surface properties. This is the first report of partitioning based on the heterogeneity of the cell surface properties correlated with the intracellular secondary metabolism in cultured plant cells. Our results also suggested that the ATPS was appropriate as a large-scale method for selecting useful cell lines among the cultured plant cells.
Edahiro
Jun-ichi
J
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Yamada
Masumi
M
Seike
Shinya
S
Kakigi
Yasuhiro
Y
Miyanaga
Kazuhiko
K
Nakamura
Masayuki
M
Kanamori
Toshiyuki
T
Seki
Minoru
M
D000872:Anthocyanins / Q000096:biosynthesis*; D002021:Buffers; D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D031985:Fragaria / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D018020:Lithium Compounds / Q000494:pharmacology; D010710:Phosphates / Q000494:pharmacology; D017680:Potassium Compounds / Q000494:pharmacology; D013431:Sulfates / Q000494:pharmacology; D014867:Water
D000872:Anthocyanins; D002021:Buffers; D018020:Lithium Compounds; D010710:Phosphates; D017680:Potassium Compounds; D013431:Sulfates; D014867:Water; C054097:lithium sulfate; C013216:potassium phosphate