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Effects of nitrogen source, calcium concentration and osmotic stress on protoplasts and protoplast‐derived cell cultures of Pinus Pinaster cotyledons
Author(s) -
David Hélène,
Jarlet Edith,
David Alain
Publication year - 1984
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1984.tb06360.x
Subject(s) - protoplast , pinus pinaster , osmotic shock , mannitol , osmotic concentration , osmotic pressure , calcium , biology , botany , cotyledon , incubation , glutamine , plant cell , plating efficiency , horticulture , biochemistry , cell , chemistry , amino acid , organic chemistry , gene
The experiments described emphasize the effects of several factors crucial to the maintenance of cell divisions leading to increased cell numbers in suspension and colony formation from cotyledon protoplasts of Pinus Pinaster Ait. Osmotic potential of the incubation and culture media are critical. Reducing the osmolality from 680 mOsm kg H 2 O −1 during protoplast isolation to 610 mOsm kg H 2 O −1 during washing and culture was essential to achieve a high frequency of cell division. Survival of the cells beyond 3 weeks of culture occurs only if the calcium concentration is decreased from 5.6 m M to 1.5 m M . Glutamine as sole source of nitrogen shortens the lag phase of response of the protoplasts and increases their plating efficiency. After 6 weeks of culture, a combination of low osmolality (225 mOsm kg H 2 O −1 ) and high level of glutamine (40 m M ) is a prerequisite for obtaining actively growing cell suspensions.