Interference of cell wall regeneration ofBoergesenia forbesii protoplasts by Tinopal LPW,a fluorescent brightening agent |
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Authors: | T Itoh R M O'Neil R M Brown Jr |
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Institution: | (1) Department of Botany, The University of Texas, 78712 Austin, TX, USA;(2) Present address: Wood Research Institute, Kyoto University, Uji, 611 Kyoto, Japan |
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Abstract: | Summary Wounding cells ofBoergesenia forbesii (Harvey) Feldmann induces the synchronous formation of numerous protoplasts which synthesize large cellulose microfibrils within 2–3 hours after wounding. The microfibrils appear to be assembled by linear terminal synthesizing complexes (TCs). TC subunits appear on both E- and P-faces of the plasma membrane, thus suggesting the occurrence of a transmembrane complex. The direction of microfibril synthesis is random during primary wall assembly and becomes ordered during secondary wall assembly. The average density of TCs during secondary wall deposition is 1.7/m2, and the average length of the TC is 510 nm. TC organization is similar to that ofValonia macrophysa; however, the larger TCs ofBoergesenia (510 nm vs. 350 nm) produce correspondingly larger microfibrils (30 nm vs. 20 nm).The effects of a fluorescent brightening agent (FBA), Tinopal LPW, on cell wall regeneration ofBoergesenia protoplasts was investigated. The threshold level of Tinopal LPW for interfering with microfibril assembly is 1.5 M. At 95 M Tinopal (for short periods up to 15 minutes), microfibril impressions have atypical spherical impressions at their termini. At longer incubations (24 hours), TCs and microfibril impressions are absent. When washed free of Tinopal, the protoplasts eventually resume normal wall assembly; however, TCs do not reappear until at least 30 minutes after the removal of Tinopal. In consideration of the presence of ordered TCs before FBA treatment, their random distribution upon recovery implies an intermediate stage of assembly or possiblyde novo synthesis. |
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Keywords: | Boergesenia forbesii Cellulose microfibrils Cell wall Fluorescent brightening agent Freeze fracture Terminal synthesizing complex |
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