Synchronization of Asparagus Somatic Embryos and Evaluation of Their Stability During Long-term Passage

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  • アスパラガス不定胚の同調化と長期継代培養時における不定胚の安定性評価
  • アスパラガス フテイハイ ノ ドウチョウカ ト チョウキ ケイダイ バイヨウジ ニ オケル フテイハイ ノ アンテイセイ ヒョウカ

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The synchronization of plant somatic embryos is a key step in achieving the stable regeneration of plantlets. In this study, we established a method for synchronizing somatic embryos induced from asparagus, Asparagus officinalis L., and evaluated the embryos from the viewpoint of morphological transition and genetic stability during long-term passage. In induction cultures of somatic embryos,the total packed cell volume (PCV) of embryos depended on the initial PCV of embryogenic calli (EC). At 21 days after induction, the PCV of obtained embryos was maximized at 43 ml/100 ml-medium when the initial PCV of EC was 1.0×10-2 ml/100 ml-medium. Mesh sieving of these somatic embryos yielded 60.6% embryo population which could develop to heart-shaped and torpedo-shaped embryos, meaning that the determined conditions were appropriate for inducing asparagus somatic embryos. Furthermore, it was demonstrated that the projected area and circularity of somatic embryos could be parameters for morphological estimation during passaging. For somatic embryos undergoing mesh sieving 42 days after induction, embryo synchronization was confirmed in terms of the projected area and circularity at 1.0-4.0 mm2 and 1.2-1.6, respectively. From morphological assessment, it was found that prolonged passaging induced abnormal embryos. Genetic analysis was conducted using plantlets regenerated from somatic embryos harvested at 42 days and 70 days after induction by means of RAPD-PCR(random amplified polymorphic DNA-polymerase chain reaction), and it was suggested that some of the embryos suffered genetic alternations through long-term passaging.

Journal

  • Shokubutsu Kankyo Kogaku

    Shokubutsu Kankyo Kogaku 20 (3), 176-183, 2008

    Japanese Society of Agricultural, Biological and Environmental Engineers and Scientists

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