Abstract
We propose a novel Madake (Phyllostachys bambusoides) bamboo suspension culture model for investigation of key enzyme(s) activity involved in growth/differentiation. Sedimented Cell Volume (SCV) and fresh weight (FW) of the suspension cultured cells reached 34% (v/v) and 8.7 g in 10 μM 2,4-dichlorophenoxyacetic acid (2,4-D)-containing medium while only 7% (v/v) SCV and 1.9 g FW of the cells in 10 μM gibberellic acid (GA3)-containing medium in 14 days. Proportion of mitotically active cells (S to G2/M phases) at a log phase was identified as 29.5% in the former cells with tiny cytoplasmic features while 5.4% in the latter cells with elongation, wall thickening, and lignification by using flow cytometry and laser scanning microscopic analysis. The total β-glucosidase (BGL) activity under the 2,4-D condition increased from 4.8 U in day 2 to 26.2 U in day 14 (ca. 5.5-fold) while a slight reduction, from 4.4 U in day 2 to 2.1 U in day 14 (ca. 0.5-fold), occurred when cell division was suppressed under the GA3 condition. Ratio of the BGL activity of the soluble fractions to the membrane-associated fractions varied depending of the culture condition. The ratio was stable (2 to 8) during the culture period under the 2,4-D condition. Interestingly, the activity of the soluble enzyme fractions increased up to ca. 65% under the GA3 condition in inverse proportion to the membrane-associated enzymes. All together, it was strongly suggested that the detected specificity/variability of BGL activity is potentially involved in cell division and lignification in Madake bamboo cells.
Highlights
The genus Phyllostachys is one of the largest genera of bamboo distributed mainly in the East Asia
We propose a novel Madake (Phyllostachys bambusoides) bamboo suspension culture model for investigation of key enzyme(s) activity involved in growth/differentiation
As the bamboo cells showed a unique feature of cellobiose degradation (Table 1), we focused on an activity of the enzyme, βglucosidase (BGL), potentially responsible for its degradation
Summary
With the aim of elucidating growth features such as cell division and wall development, we previously developed callus and suspension cell culture of Phyllostachys bamboo. As the bamboo cells showed a unique feature of cellobiose degradation (Table 1), we focused on an activity of the enzyme, βglucosidase (BGL), potentially responsible for its degradation. We established a contrastive medium condition, i.e. proliferation (modified half strength Murashige and Skoog [7] medium (m1/2MS) in the presence of 2,4-D) and lignification (m1/2MS in the presence of GA3), and identified proportion of BGL activity of the soluble and the membrane-associated enzyme fractions in our bamboo suspension cell culture model with monitoring the fre-. A β-Glucosidase Activity Potentially Involved in Cell Division and Wall Development of Phyllostachys Bamboo Suspension Cells. Technical details of bamboo suspension culture model are discussed in this study
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