Abstract
Moderately strong allelopathic activities were found in four bamboo species, Bambusa multiplex cv. Houraichiku; Phyllostachys bambusoides cv. Madake; P. nigra cv. Hachiku; Sasa kurilensis cv. Chishimazasa, which are of different classification or of different ecological distributions, using the “Sandwich Method”, which assays the dried leaves on growth of lettuce seedlings. Only small difference of activity was found among the four bamboo species. In addition, “Protoplast Co-culture Method” for assay of allelopathy in a 50 μL liquid medium using a 96 well culture plate, was applied to the suspension cultures of the four bamboo species. Protoplasts were isolated from two-week cultured suspension cells of four bamboo species using Cellulase RS and Pectolyase Y-23 in 0.6 M mannitol. At low protoplast densities of bamboo, B. multiplex and P. bambusoides stimulated the recipient lettuce growth, i.e., non-spherically cell enlargement and cell divisions observed under an inverted microscope, while protoplasts of P. nigra and S. kurilensis were less stimulatory or inhibitory. Inhibitory effect of S. kurilensis was the strongest among four bamboo species. Furthermore, highly inhibitory effects of S. kurilensis protoplasts on yellow color accumulation of lettuce protoplasts were clearly observed by analysis of a scanned digital image of a 96-well culture plate. Differences and causes of the allelopathic activities were discussed comparing with other plant species studied using the same assay methods.
Highlights
For application of bamboo allelopathy as natural herbicide in agriculture, allelopathic activities of 80 species of bamboo were investigated using in vitro bioassay method of allelopathy, “sandwich method”, using lettuce as a recipient plant [1], and of several more bamboo species were investigated [2]
Strong allelopathic activities were found in four bamboo species, Bambusa multiplex cv
At low protoplast densities of bamboo, B. multiplex and P. bambusoides stimulated the recipient lettuce growth, i.e., non-spherically cell enlargement and cell divisions observed under an inverted microscope, while protoplasts of P. nigra and S. kurilensis were less stimulatory or inhibitory
Summary
For application of bamboo allelopathy as natural herbicide in agriculture, allelopathic activities of 80 species of bamboo were investigated using in vitro bioassay method of allelopathy, “sandwich method”, using lettuce as a recipient plant [1], and of several more bamboo species were investigated [2]. A new in vitro bioassay method of allelopathy, “protoplast co-culture method” was developed [5] to contribute for finding of allelochemicals and studying mechanism(s) of allelopathy at cellular level. It measures the growth of recipient lettuce protoplasts in 50 μL liquid medium using a 96-well culture plate, at both phases of cell wall formation and cell divisions. Differences of allelopathic activities and putative allelochemicals were discussed among bamboo species and other plant species
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