ISSN:
1573-5044
Keywords:
cell culture
;
plant regeneration
;
protoplast culture
;
regeneration frequency
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract Embryogenic callus was developed from young leaves of sugarcane (Saccharum spp.hybrid, cv. CoL-54). A good embryogenic callus response was achieved using MS basal medium containing 2.0 μmol (0.5 mg l-1) picloram under dark conditions at 27±1°C. Initiation of fast growing homogeneous cell suspension cultures was achieved in MS and AA media, both supplemented with g μmol (2 mg l-1) 2,4-d and 500 mg l-1 CH. Embryogenic callus was reinitiated from embryogenic cell suspension cultures using MS medium containing 30 g l-1 sucrose, 500 mg l-1 CH and 2.26 μmol (0.5 mg l-1) 2,4-d after 4–6 weeks of culture under 16-h photoperiod conditions. Plant regeneration was achieved after about 4 weeks in MS medium lacking growth regulators but containing CH (500 mg l-1) and sucrose (60 g l-1). Rooting was enhanced by transferring regenerated plantlets to half strength MS basal medium. Totipotent protoplasts with an average yield of 2.0×107 to 1.0×108 ml-1 were obtained from embryogenic cell suspension cultures at log phase, i.e., 4–5 days after transfer to fresh media. The best growth response was achieved when protoplasts were cultured in a modifed KM8P medium at the density of 2.0×105 m l-1. Protoplasts were mainly embedded in 0.8% sea plaque agarose. Division efficiency of 22.2% was achieved after 20 days of culture and 0.26% of microcolonies continued growth and formed microcalluses after 30 days of culture under dark conditions. Microcalluses were proliferated in MS medium having 2,4-d (2 mg l-1) under 16-h photoperiod. Transferring these embryogenic calluses in MS medium +9.29 μmol kinetin (2 mg l-1) +5.37 μmol NAA (1.0 mg l-1) + activated charcoal (200 mg l-1) for 5 weeks favoured plant regeneration. Shoots and roots were further proliferated in half strength MS basal medium for 2–4 weeks. Regenerated plants were transferred to autoclaved sand for 2 weeks under 16-h photoperiod in growth room and transferred to soil in a greenhouse to raise to maturity.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00045915
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