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Effect of cellulases on spontaneous fusion of maize protoplasts

✍ Scribed by Guang-Ning Ye; Elizabeth D. Earle

Publisher: Springer
Year: 1991
Tongue: English
Weight: 613 KB
Volume: 10
Category: Article
ISSN: 0721-7714
DOI: 10.1007/bf00234298

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✦ Synopsis

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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