Induction and long-term maintenance of callus from endosperm tissue of calamansi (X Citro fortunella mitis J. Ingram and H. Moore)
1993
Patena, L.F. | Avenido, R.A. | Dimaculangan, J.G. | Barba, R.C. (Philippines Univ. Los Banos, College, Laguna (Philippines). Inst. of Plant Breeding)
Endosperm tissues were successfully excised from 60-64 day old seeds of calamansi. Primary calli were preferentially induced (33-35 percent) from the mid portion of the endosperm tissue using Murashige and Tucker (MT) medium (1966) supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg/L 6-benzylaminopurine (BAP). The primary calli proliferated when subcultured onto MT medium with 2.0 mg/L 2,4-D, 5.0 mg/L BAP and 5 percent sucrose. Callus growth was better under diffused light (75 percent) than in complete darkness (0) or under lighted condition (25 percent). Subsequent monthly subcultures of calli led to the establishment of long-term callus cultures which to this time are one year and seven months old. These long-term callus cultures proliferated and formed green compact meristemoids when subcultured onto Murashige and Skog (MS) medium with 0.5 mg/L BAP and 1 percent galactose. The aged calli (a condition required for shoot regeneration in the case of calamansi, Avenido, et al, 1991) are currently subjected to conditions for plantlet regeneration with the ultimate aim of producing plants with seedless fruits
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