Summary
The effect of protoplast-isolating enzymes on spontaneous fusion of maize protoplasts (Zea mays L. cv. Black Mexican Sweet) was investigated using a convenient ethidium bromide nuclear staining procedure. After 2–2.5 hour digestion in an enzyme solution containing 1% Cellulysin, 0.5% Rhozyme, and 0.02% Pectolyase Y-23, 50–75% of the protoplasts contained multiple nuclei. The cellulase Cellulysin was identified as the factor causing the spontaneous protoplast fusion; when Cellulysin was replaced by CELF cellulase, most protoplasts were uninucleate. Calcium and other components in the enzyme solution did not affect spontaneous fusion. Cellulysin also increased the percentage of multinucleate protoplasts from rice and asparagus suspensions. Presence of multiple nuclei might affect genetic manipulations involving protoplasts.
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References
Boss WF, Grimes HD, Brightman A (1984) Protoplasma 120:209–215
Chourey PS, Zurawski DB (1981) Theor Appl Genet 59:341–344
Fromm M, Taylor LP, Walbot V (1985) Proc Natl Acad Sci 82:5824–5828
Fromm M, Taylor LP, Walbot V (1986) Nature 319:791–793
Fromm M, Callis J, Taylor LP, Walbot V (1987) Methods Enzymol 153:351–366
Fuchs Y, Saxena A, Gamble HR, Anderson JD (1989) Plant Physiol 89:138–143
Kamo KK, Chang KL, Lynn ME, Hodges TK (1987) Planta 172:245–251
Kuang VD, Shamina ZB, Butenko RG (1984) Fiziol Rast 30:803–812
Laloue M, Courtier D, Manigault P (1980) Plant Sci Lett 17:175–179.
Ludwig SR, Somers DA, Petersen WL, Pohlman RF, Zarowitz MA, Gengenbach BG, Messing J (1985) Theor Appl Genet 71:344–350
Miller RA, Gamborg OL, Keller WA, Kao KN (1971) Can J Genet Cytol 13:347–353
Motoyoshi F (1971) Exptl Cell Res 68:452–456
Nea LJ, Bates GW (1987) Plant Cell Rep 6:337–340
Somers DA, Birnberg PR, Peterson WL, Brenner ML (1987) Plant Science 53:229–256
van der Valk HCPM, Blaas J, van Eck JW, Verhoeven HA (1988) Plant Cell Rep 7:489–492
Widholm J (1972) Stain Technol 47:189–193
Withers LA, Cocking EC (1972) J Cell Sci 11:59–75
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Communicated by G. C. Phillips
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Ye, GN., Earle, E.D. Effect of cellulases on spontaneous fusion of maize protoplasts. Plant Cell Reports 10, 213–216 (1991). https://doi.org/10.1007/BF00234298
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DOI: https://doi.org/10.1007/BF00234298