A <i>β</i>-Glucosidase Activity Potentially Involved in Cell Division and Wall Development of <i>Phyllostachys</i> Bamboo Suspension Cells

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 (GA 3 )-containing medium in 14 days. Proportion of mitotically active cells (S to G 2 /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 GA 3 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 GA 3 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.

Occurrence of hydroxycinnamoylputrescines in xylogenic bamboo suspension cells

“…After measuring the SCV, the cells were extracted and analyzed by HPLC as described above. To monitor the lignification profile of the cells, a portion of the collected cells was fixed and stained with phloroglucinol-HCl reagent (Siegel 1953) in a 96-well microplate as described previously (Ogita et al 2012c).…”

“…(madake-bamboo). Cells in these cultures proliferate in a highly synchronous manner and show uniform morphological features (Ogita 2005;Ogita et al 2012c). We also established highly efficient particle bombardment transformation protocols for suspension-cultured cells of P. nigra and P. bambusoides (Ogita et al 2011(Ogita et al , 2012a.…”

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

“…Et Zucc. (Pb, Madake) [13] [14], Bambusa multiplex Raeuschel (Bm, Houraichiku) [13] [15] and Phyllostachys nigra Munro var. Henonis (Pn, Hachiku) [13] [16] [17].…”

Multiomics-based assessment of the impact of airflow on diverse plant callus cultures

“…The calli derived from these four species were routinely subcultured by transferring four pieces of callus, each approximately 500 mg in fresh weight and 10 mm in diameter, to a new Petri dish with fresh medium every four weeks. The SR and Os callus cultures were maintained at 25°C with a 16-h light:8-h dark photoperiod, with light provided by fluorescent illumination (65 μmol m −2 s −1 ), while the bamboo callus cultures were maintained at 25°C in the dark 9 .…”