Production and cryoconservation of embryogenic cultures of mandarin and mandarin hybrids

Assays were performed to obtain embryogenic callus lines from nine mandarin and mandarin hybrid cultivars by in vitro culture of ovules collected from immature fruits six weeks after anthesis. All cultivars produced embryos and loose friable callus. The small proliferations of nucellar callus from cultured ovules were suitable to recover embryogenic callus lines by periodical subculturing to fresh medium. The embryogenic callus cultures were subjected to cryoprotection with 10% (v/v) (DMSO), freezing by slow cooling, storage in liquid nitrogen and thawing by fast warming. Whole plants from these cryopreserved cultures were recovered through embryogenesis. This revised version was published online in June 2006 with corrections to the Cover Date.