Seasonal and physiological aspects of the isolation of tobacco nicotiana tabacum cultivar xanthi nc protoplasts

Cassells, A.C.; Cocker, F.M.

Physiologia Plantarum 56(1): 69-79

1982


ISSN/ISBN: 0031-9317
DOI: 10.1111/j.1399-3054.1982.tb04901.x
Accession: 006371200

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Abstract
The variabilty of tobacco (N. tabacum L. cv. Xanthi-NC) protoplasts isolated during the winter months is affected by membrane Ca, total Ca, plant age and supplementary lighting. Feeding Ca(NO3)2 and CaCl2 helps to increase protoplast stability. Feeding is important because it increases membrane Ca, but it also has other beneficial effects which may be connected with increased NO3- and Cl- (anion) uptake and cation/anion interaction. Whatever optimal feeding regime is chosen, it must be used in conjunction with the correct harvesting practice, which depends on whether or not supplementary light is given. Plants grown without supplementary light are best harvested between 8-9 wk. If supplementary light is given, older plants (with and without supplementary Ca feeding) of .apprx. 10 wk are better for protoplast isolation. Plants grown under supplementary light have low leaf shape index (LSI) values (i.e., wider leaves) compared to plants in low light. Spring and summer Ca feeding treatments had no significant effect on increasing total or membrane Ca. There was no significant correlation between total/membrane Ca and percentage protoplast viability from days 0-5; and protoplasts isolated from plants grown in shape and fed with Ca salts were no more stable than protoplasts from control plants. Irrespective of the feed treatments given, spring and summer plants older than .apprx. 50 days and grown under shaded and normal light conditions produced poor protoplasts despite the fact that there was more Ca present in the membranes compared to protoplasts of younger plants. This decrease in percentage protoplast viability appears to be associated with changes in leaf shape. As plants age from 49-63 days, the leaf shape index (LSI) values increase (i.e., leaves are becoming narrower) and % protoplast viability decreases.

Seasonal and physiological aspects of the isolation of tobacco nicotiana tabacum cultivar xanthi nc protoplasts