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Response of Six Rice Callus Cultures to Deep-Frozen Temperatures^1,3,

Callus cultures of five mutants of rice (Oryza sativa L.), which could grow in the presence of the synthetic analog of lysine, S-aminoethyl-L-cysteine, and the nonmutant strain from which the mutants were derived were studied for growth response after freezing at several temperatures. Although the mutant lines were derived from a single line, variations were noted in response to the mixture of cryoprotectants added [10% polyethylene glycol, 8% glucose, 10% dimethylsulfoxide (PGD) in H₂O, w/v] and freezing treatments. Following freezing (and subsequent thawing), growth of callus was less at lower freezing temperatures; but all cultures grew even after freezing at —196°C. After the —196°C treatment, three lines increased three to four times the initial inoculum weight after 6-, 8-, and 10-week growth periods, compared to an increase of 14 times in 4 weeks for the cell line which grew most rapidly after thawing. Because all rice lines survived a brief period at —196°C and two lines tested, later survived —196°C for more than a year, the possibility exists for long-term, deep-frozen preservation of rice callus cultures.

Key Words: Cryoprotectants • Cryogenic (frozen) preservation • Liquid nitrogen • Oryza sativa L. • Genetic variability • Tissue culture

³ Reference by the USDA to a company and/or product is only for purposes of information and does not imply approval nor recommendation of the product to the exclusion of others which may also be suitable.

² Research chemist, biochemist, and biometrician, WRRC-ARS-USDA, Berkely, Calif.; chief (and supervisory plant physiologist) and biologist, BARC-ARS-USDA, Beltsville, Md.

Received for publication June 11, 1982.