In Vitro Bioassay of Allelopathy in Four Bamboo Species; Bambusa multiplex, Phyllostachys bambusoides, P. nigra, Sasa kurilensis, Using Sandwich Method and Protoplast Co-Culture Method with Digital Image Analysis - American Journal of Plant Sciences

AJPS > Vol.8 No.7, June 2017

In Vitro Bioassay of Allelopathy in Four Bamboo Species; Bambusa multiplex, Phyllostachys bambusoides, P. nigra, Sasa kurilensis, Using Sandwich Method and Protoplast Co-Culture Method with Digital Image Analysis ()

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DOI: 10.4236/ajps.2017.87117 1,734 Downloads 4,354 Views Citations

Author(s)

Shinjiro Ogita^1,2, Hamako Sasamoto^3,4*

Affiliation(s)

¹Biotechnology Research Center and Department of Biotechnology, Toyama Prefectural University, Imizu, Japan.
²Department of Life Sciences, Faculty of Life and Environmental Sciences, Prefectural University of Hiroshima, Shoubara, Japan.
³Faculty of Environment and Information Sciences, Yokohama National University, Yokohama, Japan.
⁴Research Institute for Integrated Science, Kanagawa University, Hiratsuka, Japan.

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.

KEYWORDS

Allelopathy, Bamboo, Bioassay, Digital Image Analysis, Protoplast Culture

Share and Cite:

Ogita, S. and Sasamoto, H. (2017) In Vitro Bioassay of Allelopathy in Four Bamboo Species; Bambusa multiplex, Phyllostachys bambusoides, P. nigra, Sasa kurilensis, Using Sandwich Method and Protoplast Co-Culture Method with Digital Image Analysis. American Journal of Plant Sciences, 8, 1699-1710. doi: 10.4236/ajps.2017.87117.

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