Strong Allelopathic Activities of Leaves and Cultured Cells of <i>Spiraea thunbergii</i> Assayed by the Protoplast Co-Culture Method with Digital Image Analysis: Evaluation of <i>Cis</i>- and <i>Trans</i>-Cinnamic Acid as Allelochemicals — Oak Academic Publishing
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Strong Allelopathic Activities of Leaves and Cultured Cells of <i>Spiraea thunbergii</i> Assayed by the Protoplast Co-Culture Method with Digital Image Analysis: Evaluation of <i>Cis</i>- and <i>Trans</i>-Cinnamic Acid as Allelochemicals
Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
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Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
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Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
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Institute of Biological Control, Faculty of Agriculture, Kyusyu University, Fukuoka, Japan
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Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
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Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
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Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
1 Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
2 Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
3 Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
4 Institute of Biological Control, Faculty of Agriculture, Kyusyu University, Fukuoka, Japan
5 Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
6 Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
7 Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
The inhibitory allelopathic activities of leaves and leaf-origin suspension cultured cells of Spiraea thunbergii , and putative allelochemicals, cis - and trans -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 S . thunbergii 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 S . thunbergii protoplasts. Effects of co-culture on the three stages of lettuce protoplast growth, i.e. , cell wall formation, cell division, and yellow pigment accumulation, were examined. Protoplasts of leaf and suspension cells of S . thunbergii strongly inhibited lettuce protoplast growth at the cell division stage (100% inhibition at 80 - 100 × 10 3 mL -1 ), but not so much at the other two stages. Both cis - and trans -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.
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