Abstract
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 × 103 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.
Highlights
As a strategy for survival, plants release allelochemicals into the surrounding environment thereby inhibiting the growth of neighboring plants that share the same habitat
Protoplasts of leaf and suspension cells of S. thunbergii strongly inhibited lettuce protoplast growth at the cell division stage (100% inhibition at 80 - 100 × 103 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
We examined the effects of plant hormones, 2,4-D and benzyladenine (BA) on the protoplast divisions in the leaves of aseptic cultured seedlings, and leaf-origin suspension culture of S. thunbergii, using 50 μl liquid medium in a 96-well culture plate
Summary
As a strategy for survival, plants release allelochemicals into the surrounding environment thereby inhibiting the growth of neighboring plants that share the same habitat. The inter-specific hybrid between S. japonica [4] has been made and anthocyanin-synthesizing genes introduced by in vitro culture to change the flower-color in S. thunbergii and S. cantoniensis [5] [6]. The second strongest inhibitory allelopathic activity of S. thunbergii and less inhibitory activity of S. cantoniensis have been observed in 168 plant species from 68 families tested using a lettuce seedling growth test [3] [7]. Leaf litters of S. thunbergii showed strong inhibitory allelopathic activity on cyanobacteria Microcystis, but not on green algae, in an algal growth test [8] [9]
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