Cryopreservation of Multiple Shoot Primordia of Endangered Plant Nymphoides indica and Regeneration of Plants from Cryopreserved Tissues

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  • UEHARA Ayumi
    Department of Life Science and Engineering, Tokyo Denki University
  • MOCHIZUKI Fuma
    Department of Life Science and Engineering, Tokyo Denki University
  • MIKUNI Tomoyo
    Graduate School of Science and Engineering, Tokyo Denki University
  • KURIYAMA Akira
    Department of Life Science and Engineering, Tokyo Denki University Graduate School of Science and Engineering, Tokyo Denki University

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Other Title
  • 絶滅危惧植物ガガブタの多芽体の超低温保存 ならびに保存組織からの植物体再生
  • ゼツメツ キグ ショクブツ ガガブタ ノ タガタイ ノ チョウテイオン ホゾン ナラビニ ホゾン ソシキ カラ ノ ショクブツタイ サイセイ

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Abstract

A technique for micropropagation and cryopreservation of an endangered plant Nymphoides indica from subcultured multiple shoot primordia is described. Excised tissues from leaves cultured on Murashige and Skoog medium (MS medium) supplemented with 3 % (w/v) sucrose, 10 μmol L-1 Thidiazuron (TDZ) and 0.1 μmol L-1 1-naphthaleneacetic acid (NAA) formed multiple shoot primordia. These shoot primordia were subcultured repeatedly at 3-week intervals. Subcultured shoot primordia were cryopreserved in liquid nitrogen. Cell clumps of multiple shoot primordia treated with 5% (v/v) dimethyl sulfoxide (DMSO) and 10% (w/v) glucose were cooled at a rate of 1°C/min to -40°C in a programmed freezer, then immersed in liquid nitrogen. Pre-culture of shoot primordia on a medium containing 5% (w/v) sucrose for 2 days enhanced survival rate of cryopreserved tissues. Plants with developed shoots and roots were obtained by transferring the survived shoot primordia to MS medium lacking growth regulators.

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