Mature leaves of Secale cereale cut into narrow strips and incubated for 18 h in a mixture of cellulase (Meicelase) and pectinase (Pectinol R10) produced quantities of protoplasts. Under the same conditions leaves of Triticum aestivum, Hordeum vulgare and Avena sativa also produce protoplasts but in
Polyribosomes in protoplasts isolated from tobacco leaves
✍ Scribed by P. Ruzicska; R. Mettrie; Y. L. Dorokhov; G. Premecz; T. Oláh; G. L. Farkas
- Publisher
- Springer-Verlag
- Year
- 1979
- Tongue
- English
- Weight
- 468 KB
- Volume
- 145
- Category
- Article
- ISSN
- 0032-0935
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✦ Synopsis
Polyribosomes (polysomes)
, active in an amino acid incorporation system in vitro, were isolated from tobacco leaf protoplasts. A comparison of polysome profiles indicated that the polysome/ monosome ratio is greatly decreased in isolated protoplasts as compared to the intact leaf. In isolated protoplasts, a marked accumulation of ribosomal subunits was also found. The division of protoplasts, as investigated in the 8-cell and callus stages, was associated with a(n) (at least) partial regeneration of polysome profiles characteristic for leaves. Plasmolysis of leaves attached to the plant had no great effect on the polysome profile. However, leaf excision per se resulted in a dramatic loss of polysomes, even when the leaf tissue was floated on water. It is concluded that the isolation of the cell from its normal environment, and not the osmotic stress and associated increase in RNase activity, is the most important factor responsible for the loss of polysomes in isolated protoplasts.
📜 SIMILAR VOLUMES
Yields of 106-108 peach mesophyll cells and protoplasts-gfw -1 were obtained depending on factors such as digesting enzymes, and leaf size. Onozuka R-10 (2%) in combination with Macerase (0.5%) was found best for protoplast isolation and mediocre for cell isolation among several enzyme combinations