. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. PROTOPLASTS IN ELECTRIC FIELDS 147 Table I Analysis of the effect of steady electric fields on growth axis ricntalion of regenerating Mougeotia protoplasts No. protoplasts Rayleigh's Critical Treatment (no. experiments) R value R value Fields (2V cm ') 661 (6) 232. 1 Fields ( cm"1) 354(3) Controls 397(10) If the calculated R is less than the critical Rons.,, then it can be said that there are no significant angular trends at P = , where n = no. protoplasts (see Ch. 2 in Zar, 1974). Micro
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. PROTOPLASTS IN ELECTRIC FIELDS 147 Table I Analysis of the effect of steady electric fields on growth axis ricntalion of regenerating Mougeotia protoplasts No. protoplasts Rayleigh's Critical Treatment (no. experiments) R value R value Fields (2V cm ') 661 (6) 232. 1 Fields ( cm"1) 354(3) Controls 397(10) If the calculated R is less than the critical Rons.,, then it can be said that there are no significant angular trends at P = , where n = no. protoplasts (see Ch. 2 in Zar, 1974). Microtubule immunofluorence Many attempts to stain microtubules in protoplasts that were either embedded in agarose or attached with mgmr' poly-L-lysine (Sigma) to one side of the field chambers were unsuccessful. Finally, protoplasts were grown in chambers as described above, except that aga- rose was omitted from the growth medium, , the pro- toplasts were suspended in 1/8 PMM during the experi- ment. After field treatment, protoplasts were stained for microtubules as described by Galway and Hardham (1986). cm) as described by Erickson and Nuccitelli (1984). Resistivity of media was 1800 (1 /8 PMM) or 1000 (1 /4 PMM) ohm cm. Field strength was measured immedi- ately and monitored throughout the experiments. Chambers were left at 20°C in a light intensity (42 n Ein- stein m : • s~') at which normal elongation of protoplasts occurred. After 16 to 20 h, the chambers were removed from the fields and the axis of elongation of the proto- plasts was recorded using a Bioquant Hipad digitizing tablet (R & M Biometrics, Nashville, Tennessee) linked to a Bioquant II software package on an Apple HE micro- computer. Results Protoplast elongation in electric fields Protoplasts elongated parallel to the direction of an ap- plied electric field with a field strength of 2 V cm"1 (Fig. 2). There was an extremely significant trend in the distri- bution of angles of elongation
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