摘要
The inhibitory allelopathic activities of leaves and leaf-origin suspension cultured cells of <i>Spiraea</i> <i>thunbergii</i>, and putative allelochemicals, <i>cis</i>- and <i>trans</i>-cinnamic acid, were investigated by the protoplast co-culture method with digital image analysis (DIA-PP method) using lettuce as a recipient. The optimal conditions of 2,4-dichlorophenoxyacetic acid (2,4-D) and benzyladenine (BA) for the cell division of <i>S</i>. <i>thunbergii</i> protoplasts were first examined using 50 μL liquid MS basal medium containing 3% sucrose, and 0.8 M mannitol in a 96-well culture plate. The hormonal condition for co-culture, 1 μM 2,4-D plus 0.1 μM BA, which was optimal for lettuce protoplast growth, was sub-optimal for <i>S</i>. <i>thunbergii</i> protoplasts. Effects of co-culture on the three stages of lettuce protoplast growth, <i>i.e.</i>, cell wall formation, cell division, and yellow pigment accumulation, were examined. Protoplasts of leaf and suspension cells of <i>S</i>. <i>thunbergii</i> strongly inhibited lettuce protoplast growth at the cell division stage (100% inhibition at 80 - 100 × 10<sup>3</sup> mL<sup>-1</sup>), but not so much at the other two stages. Both <i>cis</i>- and <i>trans</i>-cinnamic acid, showed the strongest inhibition at the cell wall formation stage, and 100% inhibition at the cell division stage at 100 μM. These results were compared with those obtained in a lettuce seedling growth test, using different allelopathic plants, and their allelochemicals were studied by the DIA-PP method.
The inhibitory allelopathic activities of leaves and leaf-origin suspension cultured cells of <i>Spiraea</i> <i>thunbergii</i>, and putative allelochemicals, <i>cis</i>- and <i>trans</i>-cinnamic acid, were investigated by the protoplast co-culture method with digital image analysis (DIA-PP method) using lettuce as a recipient. The optimal conditions of 2,4-dichlorophenoxyacetic acid (2,4-D) and benzyladenine (BA) for the cell division of <i>S</i>. <i>thunbergii</i> protoplasts were first examined using 50 μL liquid MS basal medium containing 3% sucrose, and 0.8 M mannitol in a 96-well culture plate. The hormonal condition for co-culture, 1 μM 2,4-D plus 0.1 μM BA, which was optimal for lettuce protoplast growth, was sub-optimal for <i>S</i>. <i>thunbergii</i> protoplasts. Effects of co-culture on the three stages of lettuce protoplast growth, <i>i.e.</i>, cell wall formation, cell division, and yellow pigment accumulation, were examined. Protoplasts of leaf and suspension cells of <i>S</i>. <i>thunbergii</i> strongly inhibited lettuce protoplast growth at the cell division stage (100% inhibition at 80 - 100 × 10<sup>3</sup> mL<sup>-1</sup>), but not so much at the other two stages. Both <i>cis</i>- and <i>trans</i>-cinnamic acid, showed the strongest inhibition at the cell wall formation stage, and 100% inhibition at the cell division stage at 100 μM. These results were compared with those obtained in a lettuce seedling growth test, using different allelopathic plants, and their allelochemicals were studied by the DIA-PP method.
