Production of chrysanthemum periclinal chimeras through shoot regeneration from leaf explants
Production of chrysanthemum periclinal chimeras through shoot regeneration from leaf explants
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DOI：
10.5511/plantbiotechnology.15.1127a
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发表时间：
2016-03-01
影响因子：
1.6
通讯作者：
Ohtsubo, Norihiro
中科院分区：
文献类型：
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作者：
Aida, Ryutaro;Sasaki, Katsutomo;Ohtsubo, Norihiro
周缘嵌合体在菊花等无性繁殖植物中发挥着重要作用。例如，平周嵌合体导致菊花的花色变化。在这项研究中，周嵌合体生产菊花的方法进行了检查。以野生型菊花‘太平’及其转海洋浮游生物羊舌草黄绿色荧光蛋白基因（CpYGFP）的植株为材料。将两种材料的叶外植体的切面部分附着，然后分离用于进一步培养。镶嵌愈伤组织由在外植体的分离面上形成的转基因细胞和野生型细胞组成。我们检查了来自4，120个外植体的996个再生芽，发现只有一个嵌合芽似乎显示出分生嵌合。重复腋芽伸长从mericlinal嵌合体的节点产生一个L1-荧光和一个L3-荧光嵌合植物。L1嵌合体在叶和茎的表皮细胞和毛状体中显示荧光。L3嵌合体在茎和叶的中央部分的细胞中显示荧光，并且在整个根组织中也显示荧光。综上所述，我们利用荧光蛋白转基因作为选择标记，通过叶片外植体再生获得了菊花平周嵌合体。
Periclinal chimeras play important roles in vegetatively propagated plants such as chrysanthemum (Chrysanthemum morifolium). For example, periclinal chimerism causes flower color variation in chrysanthemums. In this study, a method for periclinal chimera production in chrysanthemum was examined. A wild-type plant of chrysanthemum 'Taihei' and its transgenic plant carrying a yellowish-green fluorescent protein gene from the marine plankton Chiridius poppei (CpYGFP) were used as plant materials. The cut faces of the leaf explants of both materials were partially attached and then were detached for further culture. Mosaic calli consisted of transgenic and wild-type cells formed on the detached faces of the explants. We examined 996 regenerated shoots from 4,120 explants and found only a single chimeric shoot that appeared to show mericlinal chimerism. Repeated axillary bud elongation from the nodes of the mericlinal chimera produced one L1-fluorescent and one L3-fluorescent chimeric plant. The L1 chimera showed fluorescence in the epidermal cells and trichomes of leaf and stem. The L3 chimera showed fluorescence in the cells of the central parts of stem and leaf, as well as in the whole root tissues. In summary, we obtained chrysanthemum periclinal chimeras through regeneration from leaf explants using the fluorescent protein transgene as a selection marker.