P. Anthony et al., STRATEGIES FOR PROMOTING DIVISION OF CULTURED PLANT-PROTOPLASTS - BENEFICIAL-EFFECTS OF OXYGENATED PERFLUOROCARBON, Biotechnology techniques, 9(11), 1995, pp. 777-782
Assessments have been made of physical and chemical options, alone and
in combination, for gaseous manipulation of cassava (Manihot esculent
a Crantz.) leaf protoplast cultures. Protoplasts were cultured for 25
d in liquid medium at an initial plating density of 4 x 10(5) ml(-1) o
verlying (1) agar-solidified medium (control), (2) agar medium under a
n oxygen-enriched (10 mbar, 1 min) atmosphere, (3) agar medium supplem
ented with perfluorodecalin (Flutec(R) PP5), or (4) agar medium supple
mented with oxygenated (10 mbar, 15 min) perfluorodecalin. Similar exp
erimental treatments were also set up, with glass rods embedded in the
agar medium. The mean initial plating efficiency (IPE) of protoplasts
following culture with oxygenated perfluorodecalin without glass rods
(6.7 +/- 0.6%; n = 3) was over 2-fold greater (P < 0.05) than that of
control cultures (2.6 +/- 0.2%; n = 3). The mean IPE of protoplasts c
ultured with oxygenated perfluorodecalin in the presence of glass rods
(5.8 +/- 0.2%; n = 3) was also over 2-fold greater (P < 0.05) than co
ntrols. There was no significant difference between the IPE of protopl
asts cultured under an increased oxygen atmosphere or with oxygenated
perfluorodecalin, irrespective of the presence of glass rods.