. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 146 R. G. WHITE AND R. L. OVERALL TIME PREDIGESTION 3-7h 14-30h 30h +. MICROTUBULES Polar Array CELLULOSE MICROFIBRILS Figure 1. Organization of microtubules and cell wall micronbrils in protoplasts, of the green alga Mougeotia at intervals after protoplast isolation. (Adapted from Galway and Hardham, 1986). or 1/4 PMM (350 mMsorbitol, mAf CaCl2, 1 mM MgSO4, 15 mM MES/TRIS pH ) was layered onto the protoplast suspension, then 1/8 PMM or 1/4 PMM layered on top. Protoplasts were centrifuged for 10 min at 100 x g and tho
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 146 R. G. WHITE AND R. L. OVERALL TIME PREDIGESTION 3-7h 14-30h 30h +. MICROTUBULES Polar Array CELLULOSE MICROFIBRILS Figure 1. Organization of microtubules and cell wall micronbrils in protoplasts, of the green alga Mougeotia at intervals after protoplast isolation. (Adapted from Galway and Hardham, 1986). or 1/4 PMM (350 mMsorbitol, mAf CaCl2, 1 mM MgSO4, 15 mM MES/TRIS pH ) was layered onto the protoplast suspension, then 1/8 PMM or 1/4 PMM layered on top. Protoplasts were centrifuged for 10 min at 100 x g and those at the interface between the top and second layer were removed and rinsed 3 times with 1/8 or 1/4 PMM. To the final suspension was added sea plaque agarose (FMC Corp., Rockland, Maine) in 1/8 or 1 /4 PMM, to a final concentration of agarose. The protoplasts in PMM-agarose were cultured in field cham- bers cm deep X cm wide X 4 cm long. Application of electric fields Fields were applied via Ag/AgCl salt bridges connected via agar (1%. Bacto-Agar) tubes (47 cm long; bore or — d Figure 2. Immediately after isolation, Mougeotia protoplasts are spherical (a). After 20 h, untreated protoplasts showed random orientation of elongation (h); elongation was parallel to an applied field of 2 V cm ' (c. d, e). -. location of cathode; +. location of anode. Bar = 20 ^ Please note that these images are extracted from scanned page images that may have been digitally enhanced for readability - coloration and appearance of these illustrations may not perfectly resemble the original Marine Biological Laboratory (Woods Hole, Mass. ); Marine Biological Laboratory (Woods Hole, Mass. ). Annual report 1907/08-1952; Lillie, Frank Rattray, 1870-1947; Moore, Carl Richard, 1892-; Redfield, Alfred Clarence, 1890-1983. Woods Hole, Mass. : Marine Biological Laboratory
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