Callus formation from protoplasts of Artemisia sphaerocephala Krasch and some factors influencing protoplast division

Xu, Z.; Jia, J.

Plant cell tissue and organ culture 44(2): 129-134

1996


ISSN/ISBN: 0167-6857
DOI: 10.1007/bf00048190
Accession: 008258251

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Abstract
Round wormwood (Artemisia sphaerocephala Krasch) seeds were germinated on Murashige & Skoog (1962) medium without plant growth regulators. The hypocotyls of seedlings were sliced and cultured on M1 medium with 2,4-dichlorophenoxyacetic acid (9.05 micromolar) to induce callus. The induced calluses were subcultured on the same medium. Ten day old calluses were used to isolate protoplasts in an enzyme solution with 0.65 M mannitol. Protoplast yield strongly depended upon the state of callus cultures. Certain amount of hemicellulase could improve protoplast isolation. Purified protoplasts were cultured in modified Kao & Michayluk (1975) medium with 0.60 M mannitol as osmoticum, suggesting that protoplasts of A. sphaerocephala need a high initial osmolarity. Protoplasts generally divided evenly and the percentage of first division could reach 10%. Kinetin exhibited a positive effect on initial cell division. Furthermore, we studied the effect of protoplast density and vitamin C on sustained growth of protoplasts. After forty days, 1 mm calluses in diameter formed.

Callus formation from protoplasts of Artemisia sphaerocephala Krasch and some factors influencing protoplast division