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Method for selecting anthocyanin-producing cells by a cell sorter

K Sakamoto1, K Iida, T Koyano

  • 1Saitama Laboratory, P.C.C. Technology Inc., Japan.

Planta Medica
|June 1, 1994
PubMed
Summary
This summary is machine-generated.

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A novel fluorescein isothiocyanate (FITC) staining method distinguishes anthocyanin-producing cells. This technique enables the selection of high-anthocyanin cell lines from cultured Aralia cordata cells.

Area of Science:

  • Plant Biotechnology
  • Cell Biology
  • Biochemistry

Background:

  • Anthocyanins are valuable plant pigments with diverse applications.
  • Efficient methods for selecting high-anthocyanin-producing plant cells are crucial for biotechnological advancements.
  • Existing selection methods can be labor-intensive and may lack precision.

Purpose of the Study:

  • To develop a novel and efficient method for selecting anthocyanin-producing cells.
  • To utilize fluorescein isothiocyanate (FITC) staining for distinguishing anthocyanin-rich cells.
  • To isolate and characterize a high-anthocyanin-producing cell line from Aralia cordata.

Main Methods:

  • Developed a cell staining technique using 0.2 ppm fluorescein isothiocyanate (FITC).
  • Observed differential fluorescence under a fluorescence microscope due to FITC-anthocyanin interaction.

Related Experiment Videos

  • Applied flow cytometry based on the observed fluorescence quenching phenomenon.
  • Selected high-anthocyanin-producing protoplasts from cultured Aralia cordata cells.

    • Main Results:

    • FITC staining allowed clear visual distinction between anthocyanin-producing and non-producing cells.
    • Anthocyanin-producing cells exhibited quenched fluorescence due to light absorption at lambda max 530 nm.
    • Flow cytometry histograms effectively separated the two cell populations.
    • Sorted protoplasts showed approximately 90% anthocyanin content with 60-70% viability.
    • Selected cell line exhibited a 3-fold increase in anthocyanin concentration compared to unsorted cells.

    Conclusions:

    • The novel FITC-based staining and flow cytometry method is effective for selecting anthocyanin-producing cells.
    • This technique facilitates the isolation of high-anthocyanin-producing cell lines with improved yield.
    • The developed method holds significant potential for plant metabolic engineering and pigment production.