TITLE:
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
AUTHORS:
Shinjiro Ogita, Hamako Sasamoto
KEYWORDS:
Allelopathy, Bamboo, Bioassay, Digital Image Analysis, Protoplast Culture
JOURNAL NAME:
American Journal of Plant Sciences,
Vol.8 No.7,
June
26,
2017
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.