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GrainGenes Reference Report: PCT-44-71

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Reference	PCT-44-71
Title	Plant regeneration from embryogenic cell suspensions and protoplasts in sugarcane (Saccharum spp. hybrid cv. CoL-54)
Journal	Plant Cell Tissue and Organ Culture
Year	1996
Volume	44
Pages	71-78
Author	Aftab F Zafar Y Malik K Iqbal J
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 micromole (0.5 mg l(-1) picloram under dark conditions at 27 +/- 1 degrees C. Initiation of fast growing homogeneous cell suspension cultures was achieved in MS and AA media, both supplemented with g micromole (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 1(-1) sucrose, 500 mg l(-1) CH and 2.26 micromole (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 X 10(7) to 1.0 X 10(8) 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 modified KM8P medium at the density of 2.0 X 10(5) 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 micromole kinetin (2 mg l(-1) + 5.37. micromole 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.
Keyword	2,4-d [ Show all 31 ]

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